4-(c2-6alkoxy)-substituted chalcones as therapeutic agents

ABSTRACT

The present invention pertains to compounds of the following formula: (1) wherein: R ALK  is primary or secondary aliphatic saturated C 2-6 alkyl; each of R B2 , R B3 , R B4 , and R B5  is independently —H, —OH, or —OMe; each of R 1  and R 2  is independently: —H, optionally substituted C 1-4 alkyl, or optionally substituted C 5-20 aryl; R A3  is —H, —OH, —OC(═O)R E , —OS(═O) 2 OH, or —OP(═O)(OH) 2 ; R E  is: —H, optionally substituted C 1-6 alkyl, optionally substituted C 3-20 heterocyclyl, or optionally substituted C 5-20 aryl; or a pharmaceutically acceptable salt, solvate, amide, ester, ether, chemically protected form, or prodrug thereof. The present invention also pertains to pharmaceutical compositions comprising such compounds, and the use of such compounds and compositions, both in vitro and in vivo, for both diagnosis and treatment of, for example, proliferative conditions, such as cancer, and inflammatory conditions

This application is the U.S. national phase of international applicationPCT/GB02/04462 filed 30 Sep. 2002 which designated the U.S. and claimsbenefit of GB 0123780.9, dated 3 Oct. 2001, the entire content of whichis hereby incorporated by reference.

RELATED APPLICATION

This application is related to (and where permitted by law, claimspriority to) United Kingdom patent application number GB 0123780.9 filed3 Oct. 2001, the contents of which are incorporated herein by referencein their entirety.

TECHNICAL FIELD

This invention pertains to substituted chalcones, specificallysubstituted 1-(4-C₂₋₆alkoxyphenyl)-3-phenyl-prop-1-en-3-ones, which havetherapeutic application, for example, as potent antiproliferative agentsand antiinflammatory agents. The present invention also pertains topharmaceutical compositions comprising such compounds, and the use ofsuch compounds and compositions, both in vitro and in vivo, for bothdiagnosis and treatment of, for example, proliferative conditions, suchas cancer, and inflammatory conditions.

BACKGROUND

Many clinically successful anticancer drugs are themselves eithernatural products or have been developed from naturally occurring leadcompounds. Great interest is currently being paid to drugs isolated fromnatural resources which have already been used as a medicine. The driedwhole plant of Scutellaria barbata D. Don (Labiatae) is used inTraditional Chinese Medicine as an anti-inflammatory, an antitumouragent, and a diuretic. The α,β-unsaturated ketone,(E)-1-(4′-hydroxyphenyl)but-1-en-3-one has been isolated from this plantand found to have moderate antitumour activity (IC50 of 60 μM for K562).

Various analogues of this compounds have been examined for antitumouractivity, including one class of analogs, chalcones.

Chalcone, also known as chalkone, benzylideneacetophenone,benzalacetophenone, and phenyl styryl ketone, is1,3-diphenyl-2-propen-1-one, and has the following structure:

A number of substituted chalcones have been prepared, with one or moresubstituents on the styryl phenyl group (left, A), the acyl phenyl group(right, B), and/or the double bond carbon atoms.

A number of substituted chalcones with apparent biological activity havebeen reported.

Hall et al., 1981, describe a number of substituted chalcones which werealleged to have anti-inflammatory properties. The recited compounds areshown below (see Example 10, therein) (substituent is H unless otherwisespecified): 1 (X=OH, Z=OH, L=OH), 2 (X=OH, Y=OH, Z=OH, L=OMe), 3 (Y=OH,L=NMe₂), 4 (Y=OH, L=Cl), 5 (Y=OH, K=OEt, L=OH), 6 (Y=OH, K=C₆H₅F), 7(Y=OH, L=OH), 8 (Y=OMe, K=OMe), 9 (Y=OH, J=F), and 10 (Y=OMe, L=OH).

Eda Shoei et al., 1986, describe several substituted chalcones whichwere reported to have anti-allergic activity. Compounds 1 (X=H, Y=H), 2(X=H, Y=H), 3 (X=OH, Y=H), 4 (X=OMe, Y=H), 5 (X=OMe, Y=OMe), 6 (X=NO₂,Y=H), 7 (X=NH₂, Y=H), (see Table 1, therein) are shown below.

Berryman et al., 1995, 1997, describe a number of substituted chalconeswhich are intermediates used in the preparation of certain furanone andthiofuranone compounds reported to have activity as endothelin Iantagonists.

Some of the chalcone intermediates have a 3,4-methylenedioxy group onthe A-ring, as shown in the core structure below. See, e.g., in Berrymanet al., 1995, Examples 36, 155, 187, 191, 195, 200, 201, 205, 209, 213,217, 224, 232, 238, 242, 246, 263, 268, 280, 287, 288, 289, 298, 326,345, 352, 353, 354, 355, 357, 366, 367, 368, 369, 370, 371, 378, 380,387, 405, and 406; and additionally, in Berryman et al., 1997, Examples421, 435, and 446. Various B-ring substituents are illustrated,including: 4-hydroxy; 2-methoxy; 3-methoxy; 4-methoxy;2-allyloxy-4-methoxy; 4-isopropoxy; 2,4-dimethoxy; 3,4-dimethoxy;3,4-methylenedioxy; 3,4-methylenedioxy-5-methoxy; and 3,4-ethylenedioxy.

Although many of the chalcone intermediates have an A-ring substituentwhich is 4-methoxy, one (Example 1, page 55, in Berryman et al., 1995)has a 4-(C₂₋₆alkoxy) substituent, specifically, a 4-isopropoxysubstituent, as shown below.

Ikeda Shunichi et al., 1996, describe several substituted chalconesreported to be active as antitumour agents. Compounds 1 (X=H) (alsoreferred to herein as DMU-103), 2 (X=Me), and 3 (X=Et) (see Table 1,therein) are shown below.

Ducki et al., 1998, describe several substituted chalcones which werescreened for cytotoxic activity against the human K562 human leukemiacell line. Compounds 2a–d (X=H) and 5a–d (X=Me) (see Table 3, therein)are shown below. The X=Me compounds were found to be much more activeagainst K562 cells than the X=H compounds (see Table 3 therein).Compound 2b is also referred to herein as DMU-135.

Kharazmi et al., 1999, describe a large number of substituted chalconesalleged to be suitable for the treatment of, inter alia, inflammatoryconditions and neoplasias. See, e.g., Example 1 (pages 71–94) therein;the ring numbering scheme, shown below, is illustrated at page 132therein. None of the compounds have a 4-(C₂₋₆alkoxy) substituent or a3,4-methylenedioxy substituent (using their numbering scheme).

Potter et al., 1999, 2001 a, describe several 3,4,5-trimethoxy chalconeswhich were shown to inhibit preferentially the growth of cellsexpressing cytochrome P450 enzyme CYP1B1 as compared to cells which donot. Compounds VI (X=OMe, Y=H, Z=H, cis), VII (X=OMe, Y=H, Z=H, trans),VIII (X=OH, Y=H, Z=H), IX (X=OMe, Y=OMe, Z=H), XI (X=OMe, Y=H, Z=Me) areshown below. Compound VII was reported to be 200-fold more cytotoxic tothe cell line expressing CYP1B1 than to the parental cell line notexpressing this enzyme.

Potter et al., 2001b, describes certain substituted1-(4-methoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3one of thefollowing general formula, which have therapeutic application, and whichare potent antiproliferative agents and antiinflammatory agents.

Cushman et al., 1995, describes various stilbene derviatives, which arereported to possess utility as anticancer agents.

There is a great need for additional antiproliferative agents whichoffer one or more of the following benefits:

-   (a) improved activity.-   (b) improved selectivity (e.g., against tumour cells versus normal    cells).-   (c) low cytotoxicity as a prodrug, but yields an active drug in    vivo;-   (d) complement the activity of other treatments (e.g.,    chemotherapeutic agents);-   (e) reduced intensity of undesired side-effects;-   (f) fewer undesired side-effects;-   (g) simpler methods of administration;-   (h) reduction in required dosage amounts;-   (i) reduction in required frequency of administration;-   (j) increased ease of synthesis, purification, handling, storage,    etc.;-   (k) reduced cost of synthesis, purification, handling, storage, etc.

Thus, one aim of the present invention is the provision of compoundswhich are potent antiproliferative agents, e.g., anti-cancer agents,which offer one or more of the above benefits.

The inventors have discovered that certain sub-classes of substitutedchalcones, described herein, offer one or more of the above benefits,and additionally are surprisingly and unexpectedly more active thancorresponding known analogues.

SUMMARY OF THE INVENTION

One aspect of the invention pertains to active compounds, as describedherein, which treat a proliferative condition, such as cancer.

Another aspect of the present invention pertains to a compositioncomprising a compound as described herein and a pharmaceuticallyacceptable carrier.

Another aspect of the present invention pertains to methods ofregulating (e.g., inhibiting) cell proliferation, comprising contactinga cell with an effective amount of an active compound, as describedherein, whether in vitro or in vivo.

Another aspect of the present invention pertains to methods of treatinga proliferative condition in a subject comprising administering to saidsubject a therapeutically-effective amount of an active compound, asdescribed herein. In one preferred embodiment, the proliferativecondition is cancer.

Another aspect of the present invention pertains to an active compound,as described herein, for use in a method of treatment of the human oranimal body by therapy.

Another aspect of the present invention pertains to an active compound,as described herein, for use in a method of treatment of a proliferativecondition of the human or animal body by therapy.

Another aspect of the present invention pertains to an active compound,as described herein, for use in a method of treatment of cancer of thehuman or animal body by therapy.

Another aspect of the present invention pertains to an active compound,as described herein, for use in a method of treatment of an inflammatorycondition of the human or animal body by therapy.

Another aspect of the present invention pertains to use of an activecompound, as described herein, for the manufacture of a medicament foruse in the treatment of a proliferative condition.

Another aspect of the present invention pertains to use of an activecompound, as described herein, for the manufacture of a medicament foruse in the treatment of cancer.

Another aspect of the present invention pertains to use of an activecompound, as described herein, for the manufacture of a medicament foruse in the treatment of an inflammatory condition. In one preferredembodiment, the inflammatory condition is rheumatoid arthritis,rheumatic fever, osteoarthritis, inflammatory bowel disease, psoriasis,or bronchial asthma.

In one embodiment, the proliferative condition is characterised by cellswhich express CYP1B1.

In one embodiment, the proliferative condition is characterised by cellswhich express CYP1B1, where the corresponding normal cells do notexpress CYP1B1.

Another aspect of the present invention pertains to a compound asdescribed herein, wherein R^(A3) is —H. for use in a method of diagnosisof the human or animal body. In one preferred embodiment, the diagnosisis for the presence of tumour cells expressing the CYP1B1 enzyme.

Another aspect of the present invention pertains to the use of acompound as described herein, wherein R^(A3) is —H, for the presence ofcells (e.g., tumour cells) expressing the CYP1B1 enzyme.

Another aspect of the present invention pertains to a method ofdiagnosis of a subject for the presence of cells (e.g., tumour cells)expressing the CYP1B1 enzyme, comprising:

-   -   (a) administering to the patient a compound as described herein,        wherein R^(A3) is —H;    -   (b) determining the amount of the corresponding hydroxylated        metabolite, wherein R^(A3) is —OH, which is subsequently        produced; and,    -   (c) correlating the amount with the presence or absence of the        cells (e.g., tumour cells) in the patient.

Another aspect of the present invention pertains to a kit comprising (a)the active compound, preferably provided as a pharmaceutical compositionand in a suitable container and/or with suitable packaging; and (b)instructions for use, for example, written instructions on how toadminister the active compound.

Another aspect of the present invention pertains to compounds obtainableby a method of synthesis as described herein, or a method comprising amethod of synthesis as described herein.

Another aspect of the present invention pertains to compounds obtainedby a method of synthesis as described herein, or a method comprising amethod of synthesis as described herein.

Another aspect of the present invention pertains to novel intermediates,as described herein, which are suitable for use in the methods ofsynthesis described herein.

Another aspect of the present invention pertains to the use of suchnovel intermediates, as described herein, in the methods of synthesisdescribed herein.

As will be appreciated by one of skill in the art, features andpreferred embodiments of one aspect of the invention will also pertainto other aspects of the invention.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph of cell survivial (%) versus concentration (μM) ofcompound DMU-175, for (A) the TCDD-induced MCF-7 cell line (▪) and (B)the MCF-7 cell line (▾).

FIG. 2 is a graph of cell survivial (%) versus concentration (μM) ofcompound DMU-175, for (A) the normal breast cell line MCF-10A (∘), and(B) the advanced breast cancer cell line MDA-468 (●).

DETAILED DESCRIPTION OF THE INVENTION

Compounds

One aspect of the present invention pertains to compounds of thefollowing formula:

wherein:

-   -   R^(ALK) is primary or secondary aliphatic saturated C₂₋₆alkyl;    -   each of R^(B2), R^(B3), R^(B4), and R^(B5) is independently —H,        —OH, or —OMe;    -   each of R¹ and R² is independently —H, optionally substituted        C₁₋₄alkyl, or optionally substituted C₅₋₂₀aryl;    -   R^(A3) is —H, —OH, —OC(═O)R^(E), —OS(═O)₂OH, or —OP(═O)(OH)₂;    -   R^(E) is —H, optionally substituted C₁₋₆alkyl, optionally        substituted C₃₋₂₀heterocyclyl, or optionally substituted        C₅₋₂₀aryl;        and pharmaceutically acceptable salts, solvates, amides, esters,        ethers, chemically protected forms, and prodrugs thereof.

Note that the compounds of the present invention are all of the “E”(entgegen) or “trans” form, that is, the (optionally substituted)4-methoxy-phenyl group (styryl phenyl group) and the3,5-dimethoxybenzoyl group (acyl phenyl group) are positioned “trans”with respect to one another on the carbon-carbon double bond of theprop-1-ene backbone.

Substituent R^(ALK)

R^(ALK) is primary or secondary aliphatic saturated C₂₋₆alkyl.

The primary aliphatic saturated C₂alkyl is -Et:

The primary and secondary aliphatic saturated C₃alkyls are -nPr and-iPr:

The primary and secondary aliphatic saturated C₄alkyls are -nBu, -iBu,-sBu:

Examples of primary and secondary aliphatic saturated C₅alkyls include,but are not limited to:

Examples of primary and secondary aliphatic saturated C₅alkyls include,but are not limited to:

In one embodiment, R^(ALK) is selected from: -Et, -nPr, -iPr, -nBu,-iBu, -sBu, -nPe, and -nHex.

In one embodiment, R^(ALK) is selected from: -Et, -nPr, -nBu, -nPe, and-nHex.

Note that a reference to a particular primary or secondary aliphaticsaturated C₂₋₆alkyl includes optical isomers thereof. For example, areference to -sBu include both -sBu (R) and -sBu (S).

Substituents R^(B2), R^(B3), R^(B4), and R^(B5)

Each of R^(B2), R^(B3), R^(B4), and R^(B5) is independently —H, —OH, or—OMe.

In one embodiment, one of R^(B2), R^(B3), R^(B4), and R^(B5) is —OH or—OMe, and the others are —H (“monosubstituted”).

In one embodiment, two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OH or—OMe, and the others are —H (“disubstituted”).

In one embodiment, three of R^(B2), R^(B3), R^(B4), and R^(B5) is —OH or—OMe, and the other is —H (“trisubstituted”).

In one embodiment, each of R^(B2), R^(B3), R^(B4), and R^(B5) is —OH or—OMe (“tetrasubstituted”).

In one embodiment, one of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe,and the others are independently —H or —OH (“monomethoxy”).

In one embodiment, two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe,and the others are independently —H or —OH (“dimethoxy”).

In one embodiment, two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe,and the others are independently —H or —OH (“dimethoxy”); and the two—OMe groups are not adjacent to each other.

In one embodiment, two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe;one of the others is —OH; and the last is —H (“dimethoxy-hydroxy”).

In one embodiment, two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe;one of the others is —OH; and the last is —H (“dimethoxy-hydroxy”); andthe two —OMe groups are not adjacent to each other.

In one embodiment, three of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe,and the others are independently —H or —OH (“trimethoxy”).

In one embodiment, each of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe(“tetramethoxy”).

In one embodiment, each of R^(B2), R^(B3), R^(B4), and R^(B5) isindependently —H or —OMe.

In one embodiment, one of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe,and the others are —H (“monosubstituted, monomethoxy”).

In one embodiment, two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe,and the others are —H (“disubstituted, dimethoxy”).

In one embodiment, two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe,and the others are —H (“disubstituted, dimethoxy”); and the two —OMegroups are not adjacent to each other.

In one embodiment, three of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe,and the other is —H (“trisubstituted, trimethoxy”).

In one embodiment, the compound has one of the following formulae:

Substituents R¹ and R²

Each of R¹ and R² is independently —H, optionally substituted C₁₋₄alkyl,or optionally substituted C₅₋₂₀aryl.

In one embodiment, one of R¹ and R² is —H; and the other is —H,optionally substituted C₁₋₄alkyl, or optionally substituted C₅₋₂₀aryl.

In one embodiment, R¹ is —H; and R² is —H, optionally substitutedC₁₋₄alkyl, or optionally substituted C₅₋₂₀aryl.

In one embodiment, R² is —H; and R¹ is —H, optionally substitutedC₁₋₄alkyl, or optionally substituted C₅₋₂₀aryl.

In one embodiment, each of R¹ and R² is independently —H, -Me, or -Ph.

In one embodiment, one of R¹ and R² is —H; and the other is —H, -Me, or-Ph.

In one embodiment, R¹ is —H; and R² is —H, -Me, or -Ph.

In one embodiment, R² is —H; and R¹ is —H, -Me, or -Ph.

In one embodiment, each of R¹ and R² is independently —H or -Me.

In one embodiment, one of R¹ and R² is —H; and the other is —H or -Me.

In one embodiment, R¹ is —H; and R² is —H or -Me.

In one embodiment, R² is —H; and R¹ is —H or -Me.

In one embodiment, R¹ and R² are both —H:

In one embodiment, R¹ and R² are both —H and the compound has one of thefollowing formulae:

Substituent R^(A3)

R^(A3) is —H, —OH, —OC(═O)R^(E), —OS(═O)₂OH₁ or —OP(═O)(OH)₂, whereinR^(E) is —H, optionally substituted C₁₋₆alkyl, optionally substitutedC₃₋₂₀heterocyclyl, or optionally substituted C₅₋₂₀aryl.

In one embodiment, R^(E) is selected from:

-   —CH₃ (so that —C(═O)R^(E) is —C(═O)CH₃, acetyl);-   —CH₂CH₃ (so that —C(═O)R^(E) is —C(═O)CH₂CH₃, propionyl);-   —C(CH₃)₃ (so that —C(═O)R^(E) is —C(═O)C(CH₃)₃, pivaloyl); and-   -Ph (so that —C(═O)R^(E) is —C(═O)Ph, benzoyl).

In one embodiment, R^(A3) is —OC(═O)R^(E), —OS(═O)₂OH, or —OP(═O)(OH)₂.Such compounds may conveniently be referred to herein as “esterifiedcompounds.”

In one embodiment, R^(A3) is —H, —OH, or —OC(═O)R^(E).

In one embodiment, R^(A3) is —H or —OH.

In one embodiment, R^(A3) is —H, as shown below. Such compounds mayconveniently be referred to herein as “non-hydroxylated compounds.”

In one embodiment, R^(A3) is —H and the compound has one of thefollowing formulae:

In one embodiment, R^(A3) is —H; R¹ and R² are both —H; and the compoundhas one of the following formulae:

In one embodiment, R^(A3) is —OH, as shown below. Such compounds mayconveniently be referred to herein as “hydroxylated compounds.”

In one embodiment, R^(A3) is —OH and the compound has one of thefollowing formulae:

In one embodiment, R^(A3) is —OH; R¹ and R² are both —H; and thecompound has one of the following formulae:

Some Specific Embodiments

Some specific embodiments of the present invention are shown below.

1

DMU-174 2

DMU-175 3

DMU-176 4

DMU-184 5

DMU-185 6

DMU-186 7

DMU-190 8

DMU-191 9

DMU-192 10

DMU-187 11

DMU-188 12

DMU-189 13

DMU-401 14

DMU-417 15

DMU-408 16

DMU-420 17

DMU-424 18

DMU-432Chemical Terms

The term “carbo,” “carbyl,” “hydrocarbon” and “hydrocarbyl,” as usedherein, pertain to compounds and/or groups which have only carbon andhydrogen atoms.

The term “hetero,” as used herein, pertains to compounds and/or groupswhich have at least one heteroatom, for example, multivalent heteroatoms(which are also suitable as ring heteroatoms) such as boron, silicon,nitrogen, phosphorus, oxygen, and sulfur, and monovalent heteroatoms,such as fluorine, chlorine, bromine, and iodine.

The term “saturated,” as used herein, pertains to compounds and/orgroups which do not have any carbon-carbon double bonds or carbon-carbontriple bonds.

The term “unsaturated,” as used herein, pertains to compounds and/orgroups which have at least one carbon-carbon double bond orcarbon-carbon triple bond.

The term “aliphatic,” as used herein, pertains to compounds and/orgroups which are linear or branched, but not cyclic (also known as“acyclic” or “open-chain” groups).

The term “cyclic,” as used herein, pertains to compounds and/or groupswhich have one ring, or two or more rings (e.g., spiro, fused, bridged).

The term “ring,” as used herein, pertains to a closed ring of from 3 to10 covalently linked atoms, more preferably 3 to 8 covalently linkedatoms.

The term “aromatic ring,” as used herein, pertains to a closed ring offrom 3 to 10 covalently linked atoms, more preferably 5 to 8 covalentlylinked atoms, which ring is aromatic.

The term “heterocyclic ring,” as used herein, pertains to a closed ringof from 3 to 10 covalently linked atoms, more preferably 3 to 8covalently linked atoms, wherein at least one of the ring atoms is amultivalent ring heteroatom, for example, nitrogen, phosphorus, silicon,oxygen, and sulfur, though more commonly nitrogen, oxygen, and sulfur.

The term “alicyclic,” as used herein, pertains to compounds and/orgroups which have one ring, or two or more rings (e.g., spiro, fused,bridged), wherein said ring(s) are not aromatic.

The term “aromatic,” as used herein, pertains to compounds and/or groupswhich have one ring, or two or more rings (e.g., fused), wherein atleast one of said ring(s) is aromatic.

The term “heterocyclic,” as used herein, pertains to cyclic compoundsand/or groups which have one heterocyclic ring, or two or moreheterocyclic rings (e.g., spiro, fused, bridged), wherein said ring(s)may be alicyclic or aromatic.

The term “heteroaromatic,” as used herein, pertains to cyclic compoundsand/or groups which have one heterocyclic ring, or two or moreheterocyclic rings (e.g., fused), wherein said ring(s) is aromatic.

Substituents

The phrase “optionally substituted,” as used herein, pertains to aparent group which may be unsubstituted or which may be substituted.

Unless otherwise specified, the term “substituted,” as used herein,pertains to a parent group which bears one or more substituents. Theterm “substituent” is used herein in the conventional sense and refersto a chemical moiety which is covalently attached to, appended to, or ifappropriate, fused to, a parent group. A wide variety of substituentsare well known, and methods for their formation and introduction into avariety of parent groups are also well known.

In one preferred embodiment, the substituent(s) are independentlyselected from: halo; hydroxy; ether (e.g., C₁₋₇alkoxy); formyl; acyl(e.g., C₁₋₇alkylacyl, C₅₋₂₀arylacyl); acylhalide; carboxy; ester;acyloxy; amido; acylamido; thioamido; tetrazolyl; amino; nitro; nitroso;azido; cyano; isocyano; cyanato; isocyanato; thiocyano; isothiocyano;sulfhydryl; thioether (e.g., C₁₋₇alkylthio); sulfonic acid; sulfonate;sulfone; sulfonyloxy; sulfinyloxy; sulfamino; sulfonamino; sulfinamino;sulfamyl; sulfonamido; C₁₋₇alkyl (including, e.g., C₁₋₇haloalkyl,C₁₋₇hydroxyalkyl, C₁₋₇carboxyalkyl, C₁₋₇aminoalkyl,C₅₋₂₀aryl-C₁₋₇alkyl); C₃₋₂₀heterocyclyl; or C₅₋₂₀aryl (including, e.g.,C₅₋₂₀carboaryl, C₅₋₂₀heteroaryl, C₁₋₇alkyl-C₅₋₂₀aryl andC₅₋₂₀haloaryl)).

In one preferred embodiment, the substituent(s) are independentlyselected from:

-   —F, —Cl, —Br, and —I;-   —OH;-   —OMe, —OEt, —O(tBu), and —OCH₂Ph;-   —SH;-   —SMe, —SEt, —S(tBu), and —SCH₂Ph;-   —C(═O)H;-   —C(═O)Me, —C(═O)Et, —C(═O)(tBu), and —C(═O)Ph;-   —C(═O)OH;-   —C(═O)OMe, —C(═O)OEt, and —C(═O)O(tBu);-   —C(═O)NH₂, —C(═O)NHMe, —C(═O)NMe₂, and —C(═O)NHEt;-   —NHC(═O)Me, —NHC(═O)Et, —NHC(═O)Ph, succinimidyl, and maleimidyl;-   —NH₂, —NHMe, —NHEt, —NH(iPr), —NH(nPr), —NMe₂, —NEt₂, —N(iPr)₂,    —N(nPr)₂, —N(nBu)₂, and —N(tBu)₂;-   —CN;-   —NO₂;-   -Me, -Et, -nPr, -iPr, -nBu, -tBu;-   —CF₃, —CHF₂, —CH₂F, —CCl₃, —CBr₃, —CH₂, CH₂F, —CH₂CHF₂, and —CH₂CF₃;-   —OCF₃, —OCHF₂, —OCH₂F, —OCCl₃, —OCBr₃, —OCH₂CH₂F, —OCH₂CHF₂, and-   —OCH₂CF₃;-   —CH₂OH, —CH₂CH₂OH, and —CH(OH)CH₂OH;-   —CH₂NH₂, —CH₂CH₂NH₂, and —CH₂CH₂NMe₂; and,-   optionally substituted phenyl.

The substituents are described in more detail below.

C₁₋₇alkyl: The term “C₁₋₇alkyl,” as used herein, pertains to amonovalent moiety obtained by removing a hydrogen atom from aC₁₋₇hydrocarbon compound having from 1 to 7 carbon atoms, which may bealiphatic or alicyclic, or a combination thereof, and which may besaturated, partially unsaturated, or fully unsaturated.

Examples of (unsubstituted) saturated linear C₁₋₇alkyl groups include,but are not limited to, methyl, ethyl, n-propyl, n-butyl, and n-pentyl(amyl).

Examples of (unsubstituted) saturated branched C₁₋₇alkyl groups include,but are not limited to, iso-propyl, iso-butyl, sec-butyl, tert-butyl,and neo-pentyl.

Examples of saturated alicyclic (also carbocyclic) C₁₋₇alkyl groups(also referred to as “C₃₋₇cycloalkyl” groups) include, but are notlimited to, unsubstituted groups such as cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, and norbornane, as well as substituted groups(e.g., groups which comprise such groups), such as methylcyclopropyl,dimethylcyclopropyl, methylcyclobutyl, dimethylcyclobutyl,methylcyclopentyl, dimethylcyclopentyl, methylcyclohexyl,dimethylcyclohexyl, cyclopropylmethyl and cyclohexylmethyl.

Examples of (unsubstituted) unsaturated C₁₋₇alkyl groups which have oneor more carbon-carbon double bonds (also referred to as “C₂₋₇alkenyl”groups) include, but are not limited to, ethenyl (vinyl, —CH═CH₂),2-propenyl (allyl, —CH—CH═CH₂), isopropenyl (—C(CH₃)═CH₂), butenyl,pentenyl, and hexenyl.

Examples of (unsubstituted) unsaturated C₁₋₇alkyl groups which have oneor more carbon-carbon triple bonds (also referred to as “C₂₋₇alkynyl”groups) include, but are not limited to, ethynyl (ethinyl) and2-propynyl (propargyl).

Examples of unsaturated alicyclic (also carbocyclic) C₁₋₇alkyl groupswhich have one or more carbon-carbon double bonds (also referred to as“C₃₋₇cycloalkenyl” groups) include, but are not limited to,unsubstituted groups such as cyclopropenyl, cyclobutenyl, cyclopentenyl,and cyclohexenyl, as well as substituted groups (e.g., groups whichcomprise such groups) such as cyclopropenylmethyl andcyclohexenylmethyl.

Additional examples of substituted C₃₋₇cycloalkyl groups include, butare not limited to, those with one or more other rings fused thereto,for example, those derived from: indene (C₉), indan(2,3-dihydro-1H-indene) (C₉), tetraline (1,2,3,4-tetrahydronaphthalene(C₁₀), adamantane (C₁₀), decalin (decahydronaphthalene) (C₁₂), fluorene(C₁₃), phenalene (C₁₃). For example, 2H-inden-2-yl is a C₅cycloalkylgroup with a substituent (phenyl) fused thereto.

C₃₋₂₀heterocyclyl: The term “C₃₋₂₀heterocyclyl,” as used herein,pertains to a monovalent moiety obtained by removing a hydrogen atomfrom a ring atom of a C₃₋₂₀heterocyclic compound, said compound havingone ring, or two or more rings (e.g., spiro, fused, bridged), and havingfrom 3 to 20 ring atoms, of which from 1 to 10 are ring heteroatoms, andwherein at least one of said ring(s) is a heterocyclic ring. Preferably,each ring has from 3 to 7 ring atoms, of which from 1 to 4 are ringheteroatoms.

In this context, the prefixes (e.g., C₃₋₂₀, C₃₋₇, C₅₋₆, etc.) denote thenumber of ring atoms, or range of number of ring atoms, whether carbonatoms or heteroatoms. For example, the term “C₅₋₆heterocyclyl,” as usedherein, pertains to a heterocyclyl group having 5 or 6 ring atoms.Examples of groups of heterocyclyl groups include C₃₋₂₀heterocyclyl,C₃₋₇heterocyclyl, C₅₋₇heterocyclyl.

Examples of (non-aromatic) monocyclic heterocyclyl groups include, butare not limited to, those derived from:

-   N₁: aziridine (C₃), azetidine (C₄), pyrrolidine (tetrahydropyrrole)    (C₅), pyrroline (e.g., 3-pyrroline, 2,5-dihydropyrrole) (C₅),    2H-pyrrole or 3H-pyrrole (isopyrrole, isoazole) (C₅), piperidine    (C₆), dihydropyridine (C₆), tetrahydropyridine (C₆), azepine (C₇);-   O₁: oxirane (C₃), oxetane (C₄), oxolane (tetrahydrofuran) (C₅),    oxole (dihydrofuran) (C₅), oxane (tetrahydropyran) (C₆),    dihydropyran (C₆), pyran (C₆), oxepin (C₇);-   S₁: thiirane (C₃), thietane (C₄), thiolane (tetrahydrothiophene)    (C₅), thiane (tetrahydrothiopyran) (C₆), thiepane (C₇);-   O₂: dioxolane (C₅), dioxane (C₆), and dioxepane (C₇);-   O₃: trioxane (C₆);-   N₂: imidazolidine (C₅), pyrazolidine (diazolidine) (C₅), imidazoline    (C₅), pyrazoline (dihydropyrazole) (C₅), piperazine (C₆);-   N₁O₁: tetrahydrooxazole (C₅), dihydrooxazole (C₅),    tetrahydroisoxazole (C₅), dihydroisoxazole (C₅), morpholine (C₆),    tetrahydrooxazine (C₆), dihydrooxazine (C₆), oxazine (C₆);-   N₁S₁: thiazoline (C₅), thiazolidine (C₅), thiomorpholine (C₆);-   N₂O₁: oxadiazine (C₆);-   O₁S₁: oxathiole (C₅) and oxathiane (thioxane) (C₆); and,-   N₁O₁S₁: oxathiazine (C₆).

Examples of substituted (non-aromatic) monocyclic heterocyclyl groupsinclude saccharides, in cyclic form, for example, furanoses (C₅), suchas arabinofuranose, lyxofuranose, ribofuranose, and xylofuranse, andpyranoses (C₆), such as allopyranose, altropyranose, glucopyranose,mannopyranose, gulopyranose, idopyranose, galactopyranose, andtalopyranose.

Examples of heterocyclyl groups which are also heteroaryl groups aredescribed below with aryl groups.

C₅₋₂₀aryl: The term “C₅₋₂₀aryl,” as used herein, pertains to amonovalent moiety obtained by removing a hydrogen atom from an aromaticring atom of a C₅₋₂₀aromatic compound, said compound having one ring, ortwo or more rings (e.g., fused), and having from 5 to 20 ring atoms, andwherein at least one of said ring(s) is an aromatic ring. Preferably,each ring has from 5 to 7 ring atoms. In this context, the prefixes(e.g., C₃₋₂₀, C₅₋₇, C₅₋₆, etc.) denote the number of ring atoms, orrange of number of ring atoms, whether carbon atoms or heteroatoms. Forexample, the term “C₅₋₆aryl,” as used herein, pertains to an aryl grouphaving 5 or 6 ring atoms. Examples of groups of aryl groups includeC₃₋₂₀aryl, C₅₋₇aryl, C₅₋₆aryl.

The ring atoms may be all carbon atoms, as in “carboaryl groups” (e.g.,C₅₋₂₀carboaryl).

Examples of carboaryl groups include, but are not limited to, thosederived from benzene (i.e., phenyl) (C₆), naphthalene (C₁₀), azulene(C₁₀), anthracene (C₁₄), phenanthrene (C₁₄), naphthacene (C₁₈), andpyrene (C₁₆).

Examples of aryl groups which comprise fused rings, at least one ofwhich is an aromatic ring, include, but are not limited to, groupsderived from indene (C₉), isoindene (C₉), and fluorene (C₁₃).

Alternatively, the ring atoms may include one or more heteroatoms,including but not limited to oxygen, nitrogen, and sulfur, as in“heteroaryl groups.” In this case, the group may conveniently bereferred to as a “C₅₋₂₀heteroaryl” group, wherein “C₅₋₂₀” denotes ringatoms, whether carbon atoms or heteroatoms. Preferably, each ring hasfrom 5 to 7 ring atoms, of which from 0 to 4 are ring heteroatoms.

Examples of monocyclic heteroaryl groups include, but are not limitedto, those derived from:

-   N₁: pyrrole (azole) (C₅), pyridine (azine) (C₆);-   O₁: furan (oxole) (C₅);-   S₁: thiophene (thiole) (C₅);-   N₁O₁: oxazole (C₅), isoxazole (C₅), isoxazine (C₆);-   N₂O₁: oxadiazole (furazan) (C₅);-   N₃O₁: oxatriazole (C₅);-   N₁S₁: thiazole (C₅), isothiazole (C₅);-   N₂: imidazole (1,3-diazole) (C₅), pyrazole (1,2-diazole) (C₅),    pyridazine (1,2-diazine) (C₆), pyrimidine (1,3-diazine) (C₆) (e.g.,    cytosine, thymine, uracil), pyrazine (1,4-diazine) (C₆);-   N₃: triazole (C₅), triazine (C₆); and,-   N₄: tetrazole (C₅).    Examples of heterocyclic groups (some of which are also heteroaryl    groups) which comprise fused rings, include, but are not limited to:    -   C₉heterocyclic groups (with 2 fused rings) derived from        benzofuran (O₁), isobenzofuran (O₁), indole (N₁), isoindole        (N₁), purine (N₄) (e.g., adenine, guanine), benzimidazole (N₂),        benzoxazole (N₁O₁), benzisoxazole (N₁O₁), benzodioxole (O₂),        benzofurazan (N₂O₁), benzotriazole (N₃), benzothiofuran (S₁),        benzothiazole (N₁S₁), benzothiadiazole (N₂S);    -   C₁₀heterocyclic groups (with 2 fused rings) derived from        benzodioxan (O₂), quinoline (N₁), isoquinoline (N₁), benzoxazine        (N₁O₁), benzodiazine (N₂), pyridopyridine (N₂), quinoxaline        (N₂), quinazoline (N₂);    -   C₁₃heterocyclic groups (with 3 fused rings) derived from        carbazole (N₁), dibenzofuran (O₁), dibenzothiophene (S₁); and,    -   C₁₄heterocyclic groups (with 3 fused rings) derived from        acridine (N₁), xanthene (O₁), phenoxathiin (O₁S₁), phenazine        (N₂), phenoxazine (N₁O₁), phenothiazine (N₁S₁), thianthrene        (S₂), phenanthridine (N₁), phenanthroline (N₂), phenazine (N₂).

Heterocyclic groups (including heteroaryl groups) which have a nitrogenring atom in the form of an —NH— group may be N-substituted, that is, as—NR—. For example, pyrrole may be N-methyl substituted, to giveN-methypyrrole. Examples of N-substitutents include, but are not limitedto C₁₋₇alkyl, C₃₋₂₀heterocyclyl, C₅₋₂₀aryl, and acyl groups.

Heterocyclic groups (including heteroaryl groups) which have a nitrogenring atom in the form of an —N═ group may be substituted in the form ofan N-oxide, that is, as —N(→O)═(also denoted —N⁺(→O⁻)═). For example,quinoline may be substituted to give quinoline N-oxide; pyridine to givepyridine N-oxide; benzofurazan to give benzofurazan N-oxide (also knownas benzofuroxan).

Cyclic groups may additionally bear one or more oxo (═O) groups on ringcarbon atoms. Monocyclic examples of such groups include, but are notlimited to, those derived from:

-   C₅: cyclopentanone, cyclopentenone, cyclopentadienone;-   C₆: cyclohexanone, cyclohexenone, cyclohexadienone;-   O₁: furanone (C₅), pyrone (C₆);-   N₁: pyrrolidone (pyrrolidinone) (C₅), piperidinone (piperidone)    (C₆), piperidinedione (C₆);-   N₂: imidazolidone (imidazolidinone) (C₅), pyrazolone (pyrazolinone)    (C₅), piperazinone (C₆), piperazinedione (C₆), pyridazinone (C₆),    pyrimidinone (C₆) (e.g., cytosine), pyrimidinedione (C₆) (e.g.,    thymine, uracil), barbituric acid (C₆);-   N₁S₁: thiazolone (C₅), isothiazolone (C₅);-   N₁O₁: oxazolinone (C₅).

Polycyclic examples of such groups include, but are not limited to,those derived from:

-   C₉: indenedione;-   N₁: oxindole (C₉);-   O₁: benzopyrone (e.g., coumarin, isocoumarin, chromone) (C₁₀);-   N₁O₁: benzoxazolinone (C₉), benzoxazolinone (C₁₀);-   N₂: quinazolinedione (C₁₀);-   N₄: purinone (Cg) (e.g., guanine).

Still more examples of cyclic groups which bear one or more oxo (═O)groups on ring carbon atoms include, but are not limited to, thosederived from:

-   -   cyclic anhydrides (—C(═O)—O—C(═O)— in a ring), including but not        limited to maleic anhydride (C₅), succinic anhydride (C₅), and        glutaric anhydride (C₆);    -   cyclic carbonates (—O—C(═O)—O— in a ring), such as ethylene        carbonate (C₅) and 1,2-propylene carbonate (C₅);    -   imides (—C(═O)—NR—C(═O)— in a ring), including but not limited        to, succinimide (C₅), maleimide (C₅), phthalimide, and        glutarimide (C₆);    -   lactones (cyclic esters, —O—C(═O)— in a ring), including, but        not limited to, β-propiolactone, γ-butyrolactone,        δ-valerolactone (2-piperidone), and ε-caprolactone;    -   lactams (cyclic amides, —NR—C(═O)— in a ring), including, but        not limited to, β-propiolactam (C₄), γ-butyrolactam        (2-pyrrolidone) (C₅), δ-valerolactam (C₆), and ε-caprolactam        (C₇);    -   cyclic carbamates (—O—C(═O)—NR— in a ring), such as        2-oxazolidone (C₅);    -   cyclic ureas (—NR—C(═O)—NR— in a ring), such as 2-imidazolidone        (C₅) and pyrimidine-2,4-dione (e.g., thymine, uracil) (C₆).

The above C₁₋₇alkyl, C₃₋₂₀heterocyclyl, and C₅₋₂₀aryl groups, whetheralone or part of another substituent, may themselves optionally besubstituted with one or more groups selected from themselves and theadditional substituents listed below.

Hydrogen: —H. Note that if the substituent at a particular position ishydrogen, it may be convenient to refer to the compound as being“unsubstituted” at that position.

Halo: —F, —Cl, —Br, and −I.

Hydroxy: —OH.

Ether: —OR, wherein R is an ether substituent, for example, a C₁₋₇alkylgroup (also referred to as a C₁₋₇alkoxy group, discussed below), aC₃₋₂₀heterocyclyl group (also referred to as a C₃₋₂₀hetercyclyloxygroup), or a C₅₋₂₀aryl group (also referred to as a C₅₋₂₀aryloxy group),preferably a C₁₋₇alkyl group.

C₁₋₇alkoxy: —OR, wherein R is a C₁₋₇alkyl group. Examples of C₁₋₇alkoxygroups include, but are not limited to, —OCH₃ (methoxy), —OCH₂CH₃(ethoxy) and —OC(CH₃)₃ (tert-butoxy).

Oxo (keto, -one): ═O. Examples of cyclic compounds and/or groups having,as a substituent, an oxo group (═O) include, but are not limited to,carbocyclics such as cyclopentanone and cyclohexanone; heterocyclics,such as pyrone, pyrrolidone, pyrazolone, pyrazolinone, piperidone,piperidinedione, piperazinedione, and imidazolidone; cyclic anhydrides,including but not limited to maleic anhydride and succinic anhydride;cyclic carbonates, such as propylene carbonate; imides, including butnot limited to, succinimide and maleimide; lactones (cyclic esters,—O—C(═O)— in a ring), including, but not limited to, β-propiolactone,γ-butyrolactone, δ-valerolactone, and ε-caprolactone; and lactams(cyclic amides, —NH—C(═O)— in a ring), including, but not limited to,β-propiolactam, γ-butyrolactam, δ-valerolactam, and ε-caprolactam.

Imino (imine): ═NR, wherein R is an imino substituent, for example,hydrogen, C₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀arylgroup, preferably hydrogen or a C₁₋₇alkyl group. Examples of iminogroups include, but are not limited to, ═NH, ═NMe, —NEt, and ═NPh.

Formyl (carbaldehyde, carboxaldehyde): —C(═O)H.

Acyl (keto): —C(═O)R, wherein R is an acyl substituent, for example, aC₁₋₇alkyl group (also referred to as C₁₋₇alkylacyl or C₁₋₇alkanoyl), aC₃₋₂₀heterocyclyl group (also referred to as C₃₋₂₀heterocyclylacyl), ora C₅₋₂₀aryl group (also referred to as C₅₋₂₀arylacyl), preferably aC₁₋₇alkyl group. Examples of acyl groups include, but are not limitedto, —C(═O)CH₃ (acetyl), —C(═O)CH₂CH₃ (propionyl), —C(═O)C(CH₃)₃(butyryl), and —C(═O)Ph (benzoyl, phenone).

Acylhalide (haloformyl, halocarbonyl): —C(═O)X, wherein X is —F, —Cl,—Br, or −I, preferably —Cl, —Br, or —I.

Carboxy (carboxylic acid): —COOH.

Ester (carboxylate, carboxylic acid ester, oxycarbonyl): —C(═O)OR,wherein R is an ester substituent, for example, a C₁₋₇alkyl group, aC₃₋₂₀heterocyclyl group, or a C₅₋₂₀aryl group, preferably a C₁₋₇alkylgroup. Examples of ester groups include, but are not limited to,—C(═O)OCH₃, —C(═O)OCH₂CH₃, —C(═O)OC(CH₃)₃, and —C(═O)OPh.

Acyloxy (reverse ester): —OC(═O)R, wherein R is an acyloxy substituent,for example, a C₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group, or aC₅₋₂₀aryl group, preferably a C₁₋₇alkyl group. Examples of acyloxygroups include, but are not limited to, —OC(═O)CH₃ (acetoxy),—OC(═O)CH₂CH₃, —OC(═O)C(CH₃)₃, —OC(═O)Ph, and —OC(═O)CH₂Ph.

Amido (carbamoyl, carbamyl, aminocarbonyl, carboxamide): —C(═O)NR¹R²,wherein R¹ and R² are independently amino substituents, as defined foramino groups. Examples of amido groups include, but are not limited to,—C(═O)NH₂, —C(═O)NHCH₃, —C(═O)NH(CH₃)₂, —C(═O)NHCH₂CH₃, and—C(═O)N(CH₂CH₃)₂, as well as amido groups in which R¹ and R², togetherwith the nitrogen atom to which they are attached, form a heterocyclicstructure as in, for example, piperidinocarbonyl, morpholinocarbonyl,thiomorpholinocarbonyl, and piperazinocarbonyl.

Acylamido (acylamino): —NR¹C(═O)R², wherein R¹ is an amide substituent,for example, a C₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group, or aC₅₋₂₀aryl group, preferably a C₁₋₇alkyl group, and R² is an acylsubstituent, for example, a C₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group,or a C₅₋₂₀aryl group, preferably a C₁₋₇alkyl group. Examples ofacylamido groups include, but are not limited to, —NHC(═O)CH₃,—NHC(═O)CH₂CH₃, and —NHC(═O)Ph. R¹ and R² may together form a cyclicstructure, as in, for example, for example, succinimidyl, maleimidyl,and phthalimidyl:

Thioamido (thiocarbamyl): —C(═S)NR¹R², wherein R¹ and R² areindependently amino substituents, as defined for amino groups. Examplesof amido groups include, but are not limited to, —C(═S)NH₂, —C(═S)NHCH₃,—C(═S)NH(CH₃)₂, and —C(═S)NHCH₂CH₃.

Tetrazolyl: a five membered aromatic ring having four nitrogen atoms andone carbon atom,

Amino: —NR¹R², wherein R¹ and R² are independently amino substituents,for example, hydrogen, a C₁₋₇alkyl group (also referred to asC₁₋₇alkylamino or di-C₁₋₇alkylamino), a C₃₋₂₀heterocyclyl group, or aC₅₋₂₀aryl group, preferably H or a C₁₋₇alkyl group, or, in the case of a“cyclic” amino group, R¹ and R², taken together with the nitrogen atomto which they are attached, form a heterocyclic ring having from 4 to 8ring atoms. Examples of amino groups include, but are not limited to,—NH₂, —NHCH₃, —NHCH(CH₃)₂, —N(CH₃)₂, —N(CH₂CH₃)₂, and —NHPh.

Examples of cyclic amino groups include, but are not limited to,aziridino, azetidino, piperidino, piperazino, morpholino, andthiomorpholino.

Nitro: —NO₂.

Nitroso: —NO.

Azido: —N₃.

Cyano (nitrile, carbonitrile): —CN.

Isocyano: —NC.

Cyanato: —OCN.

Isocyanato: —NCO.

Thiocyano (thiocyanato): —SCN.

Isothiocyano (isothiocyanato): —NCS.

Sulfhydryl (thiol, mercapto): —SH.

Thioether (sulfide): —SR, wherein R is a thioether substituent, forexample, a C₁₋₇alkyl group (also referred to as a C₁₋₇alkylthio group),a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀aryl group, preferably a C₁₋₇alkylgroup. Examples of C₁₋₇alkylthio groups include, but are not limited to,—SCH₃ and —SCH₂CH₃.

Sulfonic acid (sulfo): —S(═O)₂OH.

Sulfonate (sulfonic acid ester): —S(═O)₂OR, wherein R is a sulfonatesubstituent, for example, a C₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group,or a C₅₋₂₀aryl group, preferably a C₁₋₇alkyl group. Examples ofsulfonate groups include, but are not limited to, —S(═O)₂OCH₃ and—S(═O)₂OCH₂CH₃.

Sulfone (sulfonyl): —S(═O)₂R, wherein R is a sulfone substituent, forexample, a C₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀arylgroup, preferably a C₁₋₇alkyl group. Examples of sulfone groups include,but are not limited to, —S(═O)₂CH₃ (methanesulfonyl, mesyl), —S(═O)₂CF₃,—S(═O)₂CH₂CH₃, and 4-methylphenylsulfonyl (tosyl).

Sulfonyloxy: —OS(═O)₂R, wherein R is a sulfonyloxy substituent, forexample, a C₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀arylgroup, preferably a C₁₋₇alkyl group. Examples of sulfonyloxy groupsinclude, but are not limited to, —OS(═O)₂CH₃ and —OS(═O)₂CH₂CH₃.

Sulfinyloxy: —OS(═O)R, wherein R is a sulfinyloxy substituent, forexample, a C₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀arylgroup, preferably a C₁₋₇alkyl group. Examples of sulfinyloxy groupsinclude, but are not limited to, —OS(═O)CH₃ and —OS(═O)CH₂CH₃.

Sulfamino: —NR¹S(═O)₂OH, wherein R¹ is an amino substituent, as definedfor amino groups. Examples of sulfamino groups include, but are notlimited to, —NHS(═O)₂OH and —N(CH₃)S(═O)₂OH.

Sulfonamino: —NR¹S(═O)₂R, wherein R¹ is an amino substituent, as definedfor amino groups, and R is a sulfonamino substituent, for example, aC₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀aryl group,preferably a C₁₋₇alkyl group. Examples of sulfonamino groups include,but are not limited to, —NHS(═O)₂CH₃ and —N(CH₃)S(═O)₂C₆H₅.

Sulfinamino: —NR¹S(═O)R, wherein R¹ is an amino substituent, as definedfor amino groups, and R is a sulfinamino substituent, for example, aC₁₋₇alkyl group, a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀aryl group,preferably a C₁₋₇alkyl group. Examples of sulfinamino groups include,but are not limited to, —NHS(═O)CH₃ and —N(CH₃)S(═O)C₆H₅.

Sulfamyl: —S(═O)NR¹R², wherein R¹ and R² are independently aminosubstituents, as defined for amino groups. Examples of sulfamyl groupsinclude, but are not limited to, —S(═O)NH₂, —S(═O)NH(CH₃),—S(═O)N(CH₃)₂, —S(═O)NH(CH₂CH₃), —S(═O)N(CH₂CH₃)₂, and —S(═O)NHPh.

Sulfonamido: —S(═O)₂NR¹R², wherein R¹ and R² are independently aminosubstituents, as defined for amino groups. Examples of sulfonamidogroups include, but are not limited to, —S(═O)₂NH₂, —S(═O)₂NH(CH₃),—S(═O)₂N(CH₃)₂, —S(═O)₂NH(CH₂CH₃), —S(═O)₂N(CH₂CH₃)₂, and —S(═O)₂NHPh.

As mentioned above, a C₁₋₇alkyl group may be substituted with, forexample, hydroxy (also referred to as a C₁₋₇hydroxyalkyl group),C₁₋₇alkoxy (also referred to as a C₁₋₇alkoxyalkyl group), amino (alsoreferred to as a C₁₋₇aminoalkyl group), halo (also referred to as aC₁₋₇haloalkyl group), carboxy (also referred to as a C₁₋₇carboxyalkylgroup), and C₅₋₂₀aryl (also referred to as a C₅₋₂₀aryl-C₁₋₇alkyl group).

Similarly, a C₅₋₂₀aryl group may be substituted with, for example,hydroxy (also referred to as a C₅₋₂₀hydroxyaryl group), halo (alsoreferred to as a C₅₋₂₀haloaryl group), amino (also referred to as aC₅₋₂₀aminoaryl group, e.g., as in aniline), C₁₋₇alkyl (also referred toas a C₁₋₇alkyl-C₅₋₂₀aryl group, e.g., as in toluene), and C₁₋₇alkoxy(also referred to as a C₁₋₇alkoxy-C₅₋₂₀aryl group, e.g., as in anisole).

These and other specific examples of such substituted groups are alsodiscussed below.

C₁₋₇haloalkyl group: The term “C₁₋₇haloalkyl group,” as used herein,pertains to a C₁₋₇alkyl group in which at least one hydrogen atom (e.g.,1, 2, 3) has been replaced with a halogen atom (e.g., F, Cl, Br, I). Ifmore than one hydrogen atom has been replaced with a halogen atom, thehalogen atoms may independently be the same or different. Every hydrogenatom may be replaced with a halogen atom, in which case the group mayconveniently be referred to as a C₁₋₇perhaloalkyl group.” Examples ofC₁₋₇haloalkyl groups include, but are not limited to, —CF₃, —CHF₂,—CH₂F, —CCl₃, —CBr₃, —CH₂CH₂F, —CH₂CHF₂, and —CH₂CF₃.

C₁₋₇hydroxyalkyl: The term “C₁₋₇hydroxyalkyl group,” as used herein,pertains to a C₁₋₇alkyl group in which at least one hydrogen atom hasbeen replaced with a hydroxy group. Examples of C₁₋₇hydroxyalkyl groupsinclude, but are not limited to, —CH₂OH, —CH₂CH₂OH, and —CH(OH)CH₂OH.

C₁₋₇carboxyalkyl: The term “C₁₋₇carboxyalkyl group,” as used herein,pertains to a C₁₋₇alkyl group in which at least one hydrogen atom hasbeen replaced with a carboxy group. Examples of C₁₋₇carboxyalkyl groupsinclude, but are not limited to, —CH₂COOH and —CH₂CH₂COOH.

C₁₋₇aminoalkyl: The term “C₁₋₇aminoalkyl group,” as used herein,pertains to a C₁₋₇alkyl group in which at least one hydrogen atom hasbeen replaced with an amino group. Examples of C₁₋₇aminoalkyl groupsinclude, but are not limited to, —CH₂NH₂, —CH₂CH₂NH₂, and—CH₂CH₂N(CH₃)₂.

C₁₋₇alkyl-C₅₋₂₀aryl: The term “C₁₋₇alkyl-C₅₋₂₀aryl,” as used herein,describes certain C₅₋₂₀aryl groups which have been substituted with aC₁₋₇alkyl group. Examples of such groups include, but are not limitedto, tolyl (as in toluene), xylyl (as in xylene), mesityl (as inmesitylene), styryl (as in styrene), and cumenyl (as in cumene).

C₅₋₂₀aryl-C₁₋₇alkyl: The term “C₅₋₂₀aryl-C₁₋₇alkyl,” as used herein,describers certain C₁₋₇alkyl groups which have been substituted with aC₅₋₂₀aryl group. Examples of such groups include, but are not limitedto, benzyl (phenylmethyl), tolylmethyl, phenylethyl, and triphenylmethyl(trityl).

C₅₋₂₀haloaryl: The term “C₅₋₂₀haloaryl,” as used herein, describescertain C₅₋₂₀aryl groups which have been substituted with one or morehalo groups. Examples of such groups include, but are not limited to,halophenyl (e.g., fluorophenyl, chlorophenyl, bromophenyl, oriodophenyl, whether ortho-, meta-, or para-substituted), dihalophenyl,trihalophenyl, tetrahalophenyl, and pentahalophenyl.

Bidentate Substituents

Some substituents are bidentate, that is, have two points for covalentattachment. For example, a bidentate group may be covalently bound totwo different atoms on two different groups, thereby acting as a linkertherebetween. Alternatively, a bidentate group may be covalently boundto two different atoms on the same group, thereby forming, together withthe two atoms to which it is attached (and any intervening atoms, ifpresent) a cyclic or ring structure. In this way, the bidentatesubstituent may give rise to a heterocyclic group/compound and/or anaromatic group/compound. Typically, the ring has from 3 to 8 ring atoms,which ring atoms are carbon or divalent heteroatoms (e.g., boron,silicon, nitrogen, phosphorus, oxygen, and sulfur, typically nitrogen,oxygen, and sulfur), and wherein the bonds between said ring atoms aresingle or double bonds, as permitted by the valencies of the ring atoms.Typically, the bidentate group is covalently bound to vicinal atoms,that is, adjacent atoms, in the parent group.

C₁₋₇alkylene: The term “C₁₋₇alkylene,” as used herein, pertains to abidentate moiety obtained by removing two hydrogen atoms, either bothfrom the same carbon atom, or one from each of two different carbonatoms, of a C₁₋₇hydrocarbon compound having from 1 to 7 carbon atoms,which may be aliphatic or alicyclic, or a combination thereof, and whichmay be saturated, partially unsaturated, or fully unsaturated.

Examples of linear saturated C₁₋₇alkylene groups include, but are notlimited to, —(CH₂)_(n)— where n is an integer from 1 to 7, for example,—CH₂— (methylene), —CH₂CH₂— (ethylene), —CH₂CH₂CH₂— (propylene), and—CH₂CH₂CH₂CH₂—(butylene).

Examples of branched saturated C₁₋₇alkylene groups include, but are notlimited to, —CH(CH₃)—, —CH(CH₃)CH₂—, —CH(CH₃)CH₂CH₂—,—CH(CH₃)CH₂CH₂CH₂—, —CH₂CH(CH₃)CH₂—, —CH₂CH(CH₃)CH₂CH₂—, —CH(CH₂CH₃)—,—CH(CH₂CH₃)CH₂—, and —CH₂CH(CH₂CH₃)CH₂—.

Examples of linear partially unsaturated C₁₋₇alkylene groups include,but are not limited to, —CH═CH— (vinylene), —CH═CH—CH₂—,—CH═CH—CH₂—CH₂—, —CH═CH—CH₂—CH₂—CH₂—, —CH═CH—CH═CH—, —CH═CH—CH═CH—CH₂—,—CH═CH—CH═CH—CH₂—CH₂—, —CH═CH—CH₂—CH═CH—, and —CH═CH—CH₂—CH₂—CH═CH—.

Examples of branched partially unsaturated C₁₋₇alkylene groups include,but are not limited to, —C(CH₃)═CH—, —C(CH₃)═CH—CH₂—, and—CH═CH—CH(CH₃)—.

Examples of alicyclic saturated C₁₋₇alkylene groups include, but are notlimited to, cyclopentylene (e.g., cyclopent-1,3-ylene), andcyclohexylene (e.g., cyclohex-1,4-ylene).

Examples of alicyclic partially unsaturated C₁₋₇alkylene groups include,but are not limited to, cyclopentenylene (e.g.,4-cyclopenten-1,3-ylene), cyclohexenylene (e.g., 2-cyclohexen-1,4-ylene,3-cyclohexen-1,2-ylene, 2,5-cyclohexadien-1,4-ylene).

C₅₋₂₀arylene: The term “C₅₋₂₀arylene,” as used herein, pertains to abidentate moiety obtained by removing two hydrogen atoms, one from eachof two different ring atoms of a C₅₋₂₀aromatic compound, said compoundhaving one ring, or two or more rings (e.g., fused), and having from 5to 20 ring atoms, and wherein at least one of said ring(s) is anaromatic ring. Preferably, each ring has from 5 to 7 ring atoms.

The ring atoms may be all carbon atoms, as in “carboarylene groups,” inwhich case the group may conveniently be referred to as a“C₅₋₂₀carboarylene” group.

Alternatively, the ring atoms may include one or more heteroatoms,including but not limited to oxygen, nitrogen, and sulfur, as in“heteroarylene groups.” In this case, the group may conveniently bereferred to as a “C₅₋₂₀heteroarylene” group, wherein “C₅₋₂₀” denotesring atoms, whether carbon atoms or heteroatoms. Preferably, each ringhas from 5 to 7 ring atoms, of which from 0 to 4 are ring heteroatoms.

Examples of C₅₋₂₀arylene groups which do not have ring heteroatoms(i.e., C₅₋₂₀carboarylene groups) include, but are not limited to, thosederived from benzene (i.e., phenyl) (C₆), naphthalene (C₁₀), anthracene(C₁₄), phenanthrene (C₁₄), and pyrene (C₁₆).

Examples of C₅₋₂₀heteroarylene groups include, but are not limited to,C₅heteroarylene groups derived from furan (oxole), thiophene (thiole),pyrrole (azole), imidazole (1,3-diazole), pyrazole (1,2-diazole),triazole, oxazole, isoxazole, thiazole, isothiazole, oxadiazole, andoxatriazole; and C₆heteroarylene groups derived from isoxazine, pyridine(azine), pyridazine (1,2-diazine), pyrimidine (1,3-diazine; e.g.,cytosine, thymine, uracil), pyrazine (1,4-diazine), triazine,tetra-zole, and oxadiazole (furazan).

C₅₋₂₀Arylene-C₁₋₇alkylene: The term “C₅₋₂₀arylene-C₁₋₇alkylene,” as usedherein, pertains to a bidentate moiety comprising a C₅₋₂₀arylene moiety,-Arylene-, linked to a C₁₋₇alkylene moiety, -Alkylene-, that is,-Arylene-Alkylene-.

Examples of C₅₋₂₀arylene-C₁₋₇alkylene groups include, but are notlimited to, phenylene-methylene, phenylene-ethylene,phenylene-propylene, and phenylene-ethenylene (also known asphenylene-vinylene).

C₅₋₂₀Alkylene-C₁₋₇arylene: The term “C₅₋₂₀alkylene-C₁₋₇arylene,” as usedherein, pertains to a bidentate moiety comprising a C₅₋₂₀alkylenemoiety, -Alkylene-, linked to a C₁₋₇arylene moiety, -Arylene-, that is,-Alkylene-Arylene-.

Examples of C₅₋₂₀alkylene-C₁₋₇arylene groups include, but are notlimited to, methylene-phenylene, ethylene-phenylene,propylene-phenylene, and ethenylene-phenylene (also known asvinylene-phenylene).

Included in the above are the well known ionic, salt, solvate (e.g.,hydrate), and protected forms of these substituents. For example, areference to carboxylic acid (—COOH) also includes carboxylate (—COO⁻).Similarly, a reference to an amino group includes a salt, for example, ahydrochloride salt, of the amino group. A reference to a hydroxyl groupalso includes conventional protected forms of a hydroxyl group.Similarly, a reference to an amino group also includes conventionalprotected forms of an amino group.

Acronyms

For convenience, many chemical moieties are represented herein usingwell known abbreviations, including but not limited to, methyl (Me),ethyl (Et), n-propyl (nPr), iso-propyl (iPr), n-butyl (nBu), tert-butyl(tBu), n-hexyl (nHex), cyclohexyl (cHex), phenyl(Ph), biphenyl (biPh),benzyl (Bn), naphthyl (naph), methoxy (MeO), ethoxy (EtO), benzoyl (Bz),and acetyl (Ac).

For convenience, many chemical compounds are represented herein usingwell known abbreviations, including but not limited to, methanol (MeOH),ethanol (EtOH), iso-propanol (i-PrOH), methyl ethyl ketone (MEK), aceticacid (AcOH), dichloromethane (methylene chloride, DCM), trifluoroaceticacid (TFA), dimethylformamide (DMF), and tetrahydrofuran (THF).

Isomers, Salts, Solvates, Protected Forms, and Prodrugs

A certain compound may exist in one or more particular geometric,optical, enantiomeric, diasteriomeric, epimeric, stereoisomeric,tautomeric, conformational, or anomeric forms, including but not limitedto, cis- and trans-forms; E- and Z-forms; c-, t-, and r-forms; endo- andexo-forms; R-, S-, and meso-forms; D- and L-forms; (+) and (−) forms;keto-, enol-, and enolate-forms; syn- and anti-forms; synclinal- andanticlinal-forms; α- and β-forms; axial and equatorial forms; boat-,chair-, twist-, envelope-, and halfchair-forms; and combinationsthereof, hereinafter collectively referred to as “isomers” (or “isomericforms”).

Note that, except as discussed below for tautomeric forms, specificallyexcluded from the term “isomers,” as used herein, are structural (orconstitutional) isomers (i.e., isomers which differ in the connectionsbetween atoms rather than merely by the position of atoms in space). Forexample, a reference to a methoxy group, —OCH₃, is not to be construedas a reference to its structural isomer, a hydroxymethyl group, —CH₂OH.Similarly, a reference to ortho-chlorophenyl is not to be construed as areference to its structural isomer, meta-chlorophenyl. However, areference to a class of structures may well include structurallyisomeric forms falling within that class (e.g., C₁₋₇alkyl includesn-propyl and iso-propyl; butyl includes n-, iso-, sec-, and tert-butyl;methoxyphenyl includes ortho-, meta-, and para-methoxyphenyl).

The above exclusion does not pertain to tautomeric forms, for example,keto-, enol-, and enolate-forms, as in, for example, the followingtautomeric pairs: keto/enol (illustrated below), imine/enamine,amide/imino alcohol, amidine/amidine, nitroso/oxime,thioketone/enethiol, N-nitroso/hydroxyazo, and nitro/aci-nitro.

Note that specifically included in the term “isomer” are compounds withone or more isotopic substitutions. For example, H may be in anyisotopic form, including ¹H, ²H (D), and ³H (T); C may be in anyisotopic form, including ¹²C, ¹³C, and ¹⁴C; O may be in any isotopicform, including ¹⁶O and ¹⁸O; and the like.

Unless otherwise specified, a reference to a particular compoundincludes all such isomeric forms, including racemic and other mixturesthereof. Methods for the preparation (e.g., asymmetric synthesis) andseparation (e.g., fractional crystallisation and chromatographic means)of such isomeric forms are either known in the art or are readilyobtained by adapting the methods taught herein in a known manner.

Unless otherwise specified, a reference to a particular compound alsoincludes ionic, salt, solvate (e.g., hydrate), protected forms, andprodrugs thereof, for example, as discussed below.

It may be convenient or desirable to prepare, purify, and/or handle acorresponding salt of the active compound, for example, apharmaceutically-acceptable salt. Examples of pharmaceuticallyacceptable salts are discussed in Berge et al., 1977, “PharmaceuticallyAcceptable Salts,” J. Pharm. Sci., Vol. 66, pp. 1–19.

For example, if the compound is anionic, or has a functional group whichmay be anionic (e.g., —COOH may be —COO⁻), then a salt may be formedwith a suitable cation. Examples of suitable inorganic cations include,but are not limited to, alkali metal ions such as Na+ and K+, alkalineearth cations such as Ca²⁺ and Mg²⁺, and other cations such as Al⁺³.Examples of suitable organic cations include, but are not limited to,ammonium ion (i.e., NH₄ ⁺) and substituted ammonium ions (e.g., NH₃R⁺,NH₂R₂ ⁺, NHR₃ ⁺, NR₄ ⁺). Examples of some suitable substituted ammoniumions are those derived from: ethylamine, diethylamine, ethylenediamine,ethanolamine, diethanolamine, piperazine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

If the compound is cationic, or has a functional group which may becationic (e.g., —NH₂ may be —NH₃ ⁺), then a salt may be formed with asuitable anion. Examples of suitable inorganic anions include, but arenot limited to, those derived from the following inorganic acids:hydrochloric, hydrobromic, hydroiodic, sulfuric, sulfurous, nitric,nitrous, phosphoric, and phosphorous. Examples of suitable organicanions include, but are not limited to, anions from the followingorganic acids: acetic, propionic, succinic, gycolic, stearic, lactic,malic, tartaric, citric, ascorbic, maleic, hydroxymaleic, phenylacetic,glutamic, benzoic, salicylic, sulfanilic, 2-acetyoxybenzoic, fumaric,toluenesulfonic, methanesulfonic, ethanesulfonic, ethane disulfonic,oxalic, isethionic, and valeric.

It may be convenient or desirable to prepare, purify, and/or handle acorresponding solvate of the active compound. The term “solvate” is usedherein in the conventional sense to refer to a complex of solute (e.g.,active compound, salt of active compound) and solvent. If the solvent iswater, the solvate may be conveniently referred to as a hydrate, forexample, a mono-hydrate, a di-hydrate, a tri-hydrate, etc.

It may be convenient or desirable to prepare, purify, and/or handle theactive compound in a chemically protected form. The term “chemicallyprotected form,” as used herein, pertains to a compound in which one ormore reactive functional groups are protected from undesirable chemicalreactions, that is, are in the form of a protected or protecting group(also known as a masked or masking group). By protecting a reactivefunctional group, reactions involving other unprotected reactivefunctional groups can be performed, without affecting the protectedgroup; the protecting group may be removed, usually in a subsequentstep, without substantially affecting the remainder of the molecule.See, for example, Protective Groups in Organic Synthesis (T. Green andP. Wuts, Wiley, 1991), and Protective Groups in Organic Synthesis (T.Green and P. Wuts; 3rd Edition; John Wiley and Sons, 1999).

For example, a hydroxy group may be protected as an ether (—OR) or anester (—OC(═O)R), for example, as: a t-butyl ether; a benzyl, benzhydryl(diphenylmethyl), or trityl (triphenylmethyl) ether; a trimethylsilyl ort-butyldimethylsilyl ether; or an acetyl ester (—OC(═O)CH₃, —OAc).

For example, an aldehyde or ketone group may be protected as an acetalor ketal, respectively, in which the carbonyl group (>C═O) is convertedto a diether (>C(OR)₂), by reaction with, for example, a primaryalcohol. The aldehyde or ketone group is readily regenerated byhydrolysis using a large excess of water in the presence of acid.

For example, an amine group may be protected, for example, as an amide(—NRCO—R) or a urethane (—NRCO—OR), for example, as: a methyl amide(—NHCO—CH₃); a benzyloxy amide (—NHCO—OCH₂C₆H₅, —NH-Cbz); as a t-butoxyamide (—NHCO—OC(CH₃)₃, —NH-Boc); a 2-biphenyl-2-propoxy amide(—NHCO—OC(CH₃)₂C₆H₄C₆H₅, —NH-Bpoc), as a 9-fluorenylmethoxy amide(—NH-Fmoc), as a 6-nitroveratryloxy amide (—NH-Nvoc), as a2-trimethylsilylethyloxy amide (—NH-Teoc), as a 2,2,2-trichloroethyloxyamide (—NH-Troc), as an allyloxy amide (—NH-Alloc), as a2(-phenylsulfonyl)ethyloxy amide (—NH-Psec); or, in suitable cases(e.g., cyclic amines), as a nitroxide radical (>N—O*).

For example, a carboxylic acid group may be protected as an ester or anamide, for example, as: a benzyl ester; a t-butyl ester; a methyl ester;or a methyl amide.

For example, a thiol group may be protected as a thioether (—SR), forexample, as: a benzyl thioether; an acetamidomethyl ether(—S—CH₂NHC(═O)CH₃).

It may be convenient or desirable to prepare, purify, and/or handle theactive compound in the form of a prodrug. The term “prodrug,” as usedherein, pertains to a compound which, when metabolised, yields thedesired active compound. Typically, the prodrug is inactive, or lessactive than the active compound, but may provide advantageous handling,administration, or metabolic properties. For example, some prodrugs areesters of the active compound; during metabolysis, the ester group iscleaved to yield the active drug. Also, some prodrugs are activatedenzymatically to yield the active compound, or a compound which, uponfurther chemical reaction, yields the active compound. For example, theprodrug may be a sugar derivative or other glycoside conjugate, or maybe an amino acid ester derivative.

Synthesis

Several methods for the chemical synthesis of compounds of the presentinvention are described herein. These methods may be modified and/oradapted in known ways in order to facilitate the synthesis of additionalcompounds within the scope of the present invention.

The compounds of the present invention may be prepared, for example, byAldol condensation of the corresponding carbonyl compounds A and B, asillustrated below in Scheme 1.

When R¹ is —H, the first compound is a 4-alkoxybenzaldehyde. When R² is—H, the second compound is an acetophenone.

Many suitable starting reagents are commercially available (e.g., fromSigma-Aldrich). Additional reagents may be synthesised using knownmethods, or by modifying known methods in known ways.

For example, compound DMU-174 may be prepared by stirring a mixture of4-ethoxybenzaldehyde (A) and 3,5-dimethoxyacetophenone (B) in a suitablesolvent, e.g., methanol, with added base catalyst, e.g., aqueous sodiumhydroxide for 2 hours at ambient temperature. The reaction isillustrated below in Scheme 2.

Compounds for which R^(A3) is —OC(═O)R^(E), —OS(═O)₂OH, or —OP(═O)(OH)₂may be prepared from their hydroxy analogs (where R^(A3) is —OH) byreaction with an organic acid (i.e., R^(E)COOH) or an inorganic acid(i.e., sulfuric acid, H₂SO₄; phosphoric acid, H₃PO₄).

The groups —OS(═O)₂OH and —OP(═O)(OH)₂ may be present as such, in theirfree acid form, or they may be present as a salt or ester thereof, asdiscussed above. For example, the group —OS(═O)₂OH may be present as—OS(═O)₂O⁻ M⁺, wherein M⁺ is a suitable cation. Similarly, the group—OP(═O)(OH)₂ may be present as —OP(═O)(OH)O⁻ M⁺ or —OP(═O)(O⁻)₂(M⁺)₂,wherein M⁺ is a suitable cation. Examples of suitable cations arediscussed above. In one embodiment, the group —OP(═O)(OH)₂ is present asthe disodium salt, —OP(═O)(O⁻)₂(Na⁺)₂. Other salts and esters aredescribed in Pettit et al, 1995.

Uses

The present invention provides active compounds which are capable ofregulating (e.g., inhibiting) cell proliferation, as well as methods ofregulating (e.g., inhibiting) cell proliferation, comprising contactinga cell with an effective amount of an active compound, whether in vitroor in vivo.

The term “active,” as used herein, pertains to compounds which arecapable of regulating (e.g., inhibiting) cell proliferation, andspecifically includes both compounds with intrinsic activity (drugs) aswell as prodrugs of such compounds, which prodrugs may themselvesexhibit little or no intrinsic activity.

One of ordinary skill in the art is readily able to determine whether ornot a candidate compound is active, that is, capable of regulating(e.g., inhibiting) cell proliferation. For example, assays which mayconveniently be used to assess the proliferation regulation offered by aparticular compound are described in the examples below.

For example, a sample of cells (e.g., from a tumour) may be grown invitro and a candidate compound brought into contact with the cells, andthe effect of the compound on those cells observed. As examples of“effect,” the morphological status of the cells may be determined (e.g.,alive or dead). Where the candidate compound is found to exert aninfluence on the cells, this may be used as a prognostic or diagnosticmarker of the efficacy of the compound in methods of treating a patientcarrying cells of the same type (e.g., the tumour or a tumour of thesame cellular type).

In one aspect, the present invention provides antiproliferative agents.The term “antiproliferative agent” as used herein, pertains to acompound which treats a proliferative condition (i.e., a compound whichis useful in the treatment of a proliferative condition).

The terms “cell proliferation,” “proliferative condition,”“proliferative disorder,” and “proliferative disease,” are usedinterchangeably herein and pertain to an unwanted or uncontrolledcellular proliferation of excessive or abnormal cells which isundesired, such as, neoplastic or hyperplastic growth, whether in vitroor in vivo. Examples of proliferative conditions include, but are notlimited to, pre-malignant and malignant cellular proliferation,including but not limited to, malignant neoplasms and tumours, cancers,leukemias, psoriasis, bone diseases, fibroproliferative disorders (e.g.,of connective tissues), and atherosclerosis. Any type of cell may betreated, including but not limited to, lung, colon, breast, ovarian,prostate, liver, pancreas, brain, and skin.

In another embodiment, the proliferative condition is a solid tumour. Inanother embodiment, the proliferative condition is a solid tumour, andis a cancer of the lung, colon, breast, ovarian, prostate, liver,pancreas, brain, or skin. In another embodiment, the proliferativecondition is a solid tumour, and is a cancer of the breast.

As discussed below (see “Prodrugs”), compounds of the present inventionmay act as prodrugs useful as antiproliferative agents with lowintrinsic toxicity, for treatment of proliferative conditions which arecharacterised by cells which express the CYP1B1 enzyme.

Additionally, compounds of the present invention may act as prodrugsuseful as selective antiproliferative agents with low intrinsictoxicity, for treatment of proliferative conditions which arecharacterised by cells which express the CYP1B1 enzyme, where thecorresponding normal cells do not express the CYP1B1 enzyme.

Thus, in one preferred embodiment, the proliferative condition ischaracterised by cells which express CYP1B1. In one preferredembodiment, the proliferative condition is characterised by cells whichexpress CYP1B1, where the corresponding normal cells do not expressCYP1B1. For example, the proliferative condition may be a tumourcharacterised by tumour cells which express CYP1B1, where thecorresponding normal cells do not.

Antiproliferative compounds of the present invention have application inthe treatment of cancer, and so the present invention further providesanticancer agents. The term “anticancer agent” as used herein, pertainsto a compound which treats a cancer (i.e., a compound which is useful inthe treatment of a cancer). The anti-cancer effect may arise through oneor more mechanisms, including but not limited to, the regulation of cellproliferation, the inhibition of angiogenesis (the formation of newblood vessels), the inhibition of metastasis (the spread of a tumourfrom its origin), the inhibition of invasion (the spread of tumour cellsinto neighbouring normal structures), or the promotion of apoptosis(programmed cell death).

The present invention also provides active compounds which are useful inthe treatment of inflammatory conditions. For example, such compoundshave growth down-regulatory effects on splenocytes. Examples ofinflammaotry conditions include, but are not limited to, rheumatoidarthritis, rheumatic fever, osteoarthritis, inflammatory bowel disease,psoriasis, and bronchial asthma.

The invention further provides active compounds for use in a method oftreatment of the human or animal body by therapy. Such a method maycomprise administering to such a subject a therapeutically-effectiveamount of an active compound, preferably in the form of a pharmaceuticalcomposition.

The term “treatment,” as used herein in the context of treating acondition, pertains generally to treatment and therapy, whether of ahuman or an animal (e.g., in veterinary applications), in which somedesired therapeutic effect is achieved, for example, the inhibition ofthe progress of the condition, and includes a reduction in the rate ofprogress, a halt in the rate of progress, amelioration of the condition,and cure of the condition. Treatment as a prophylactic measure is alsoincluded.

The term “therapeutically-effective amount,” as used herein, pertains tothat amount of an active compound, or a material, composition or dosagefrom comprising an active compound, which is effective for producingsome desired therapeutic effect, commensurate with a reasonablebenefit/risk ratio.

The term “treatment” includes combination treatments and therapies, inwhich two or more treatments or therapies are combined, for example,sequentially or simultaneously. Examples of treatments and therapiesinclude, but are not limited to, chemotherapy (the administration ofactive agents, including, e.g., drugs, antibodies (e.g., as inimmunotherapy), prodrugs (e.g., as in photodynamic therapy, GDEPT,ADEPT, etc.); surgery; radiation therapy; and gene therapy.

The invention further provides the use of an active compound for themanufacture of a medicament, for example, for the treatment of aproliferative condition or an inflammatory condition, as discussedabove.

The invention further provides a method for regulating (e.g.,inhibiting) cell proliferation, comprising said cell with an effectiveamount of an active compound whether in vitro or in vivo.

Another aspect of the present invention pertains to methods of treatinga proliferative condition in a subject comprising administering to saidsubject a therapeutically-effective amount of an active compound,preferably in the form of a pharmaceutical composition.

Active compounds may also be used, as described above, in combinationtherapies, that is, in conjunction with other agents, for example,cytotoxic agents.

Active compounds may also be used as part of an in vitro assay, forexample, in order to determine whether a candidate host is likely tobenefit from treatment with the compound in question.

Active compounds may also be used as a standard, for example, in anassay, in order to identify other active compounds, otherantiproliferative agents, other antiinflammatory agents, etc.

Routes of Administration

The active compound or pharmaceutical composition comprising the activecompound may be administered to a subject by any convenient route ofadministration, whether systemically/peripherally or topically (i.e., atthe site of desired action).

Routes of administration include, but are not limited to, oral (e.g, byingestion); buccal; sublingual; transdermal (including, e.g., by apatch, plaster, etc.); transmucosal (including, e.g., by a patch,plaster, etc.); intranasal (e.g., by nasal spray); ocular (e.g., byeyedrops); pulmonary (e.g., by inhalation or insufflation therapy using,e.g., via an aerosol, e.g., through the mouth or nose); rectal (e.g., bysuppository or enema); vaginal (e.g., by pessary); parenteral, forexample, by injection, including subcutaneous, intradermal,intramuscular, intravenous, intraarterial, intracardiac, intrathecal,intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal,intratracheal, subcuticular, intraarticular, subarachnoid, andintrasternal; by implant of a depot or reservoir, for example,subcutaneously or intramuscularly.

The Subject

The subject may be a prokaryote (e.g., bacteria) or a eukaryote (e.g.,protoctista, fungi, plants, animals).

The subject may be a protoctista, an alga, or a protozoan.

The subject may be a plant, an angiosperm, a dicotyledon, amonocotyledon, a gymnosperm, a conifer, a ginkgo, a cycad, a fern, ahorsetail, a clubmoss, a liverwort, or a moss.

The subject may be an animal.

The subject may be a chordate, an invertebrate, an echinoderm (e.g.,starfish, sea urchins, brittlestars), an arthropod, an annelid(segmented worms) (e.g., earthworms, lugworms, leeches), a mollusk(cephalopods (e.g., squids, octopi), pelecypods (e.g., oysters, mussels,clams), gastropods (e.g., snails, slugs)), a nematode (round worms), aplatyhelminthes (flatworms) (e.g., planarians, flukes, tapeworms), acnidaria (e.g., jelly fish, sea anemones, corals), or a porifera (e.g.,sponges).

The subject may be an arthropod, an insect (e.g., beetles, butterflies,moths), a chilopoda (centipedes), a diplopoda (millipedes), a crustacean(e.g., shrimps, crabs, lobsters), or an arachnid (e.g., spiders,scorpions, mites).

The subject may be a chordate, a vertebrate, a mammal, a bird, a reptile(e.g., snakes, lizards, crocodiles), an amphibian (e.g., frogs, toads),a bony fish (e.g., salmon, plaice, eel, lungfish), a cartilaginous fish(e.g., sharks, rays), or a jawless fish (e.g., lampreys, hagfish).

The subject may be a mammal, a placental mammal, a marsupial (e.g.,kangaroo, wombat), a monotreme (e.g., duckbilled platypus), a rodent(e.g., a guinea pig, a hamster, a rat, a mouse), murine (e.g., a mouse),a lagomorph (e.g., a rabbit), avian (e.g., a bird), canine (e.g., adog), feline (e.g., a cat), equine (e.g., a horse), porcine (e.g., apig), ovine (e.g., a sheep), bovine (e.g., a cow), a primate, simian(e.g., a monkey or ape), a monkey (e.g., marmoset, baboon), an ape(e.g., gorilla, chimpanzee, orangutang, gibbon), or a human.

Furthermore, the subject may be any of its forms of development, forexample, a spore, a seed, an egg, a larva, a pupa, or a foetus.

In one preferred embodiment, the subject is a human.

Formulations

While it is possible for the active ingredient to be administered alone,it is preferable to present it as a pharmaceutical composition (e.g.,formulation) comprising at least one active ingredient, as definedabove, together with one or more pharmaceutically acceptable carriers,excipients, buffers, adjuvants, stabilisers, or other materials wellknown to those skilled in the art and optionally other therapeuticagents.

Thus, the present invention further provides pharmaceuticalcompositions, as defined above, and methods of making a pharmaceuticalcomposition comprising admixing at least one active ingredient, asdefined above, together with one or more pharmaceutically acceptablecarriers, excipients, buffers, adjuvants, stabilisers, or othermaterials, as described herein.

The term “pharmaceutically acceptable” as used herein pertains tocompounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgement, suitable for use in contactwith the tissues of a subject (e.g., human) without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio. Each carrier,excipient, etc. must also be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation.

The formulations may conveniently be presented in unit dosage form andmay be prepared by any methods well known in the art of pharmacy. Suchmethods include the step of bringing into association the activeingredient with the carrier which constitutes one or more accessoryingredients. In general, the formulations are prepared by uniformly andintimately bringing into association the active ingredient with liquidcarriers or finely divided solid carriers or both, and then if necessaryshaping the product.

Formulations may be in the form of liquids, solutions, suspensions,emulsions, tablets, losenges, granules, powders, capsules, cachets,pills, ampoules, suppositories, pessaries, ointments, gels, pastes,creams, sprays, foams, lotions, oils, boluses, electuaries, or aerosols.

Formulations suitable for oral administration (e.g., by ingestion) maybe presented as discrete units such as capsules, cachets or tablets,each containing a predetermined amount of the active ingredient; as apowder or granules; as a solution or suspension in an aqueous ornon-aqueous liquid; or as an oil-in-water liquid emulsion or awater-in-oil liquid emulsion; as a bolus; as an electuary; or as apaste.

A tablet may be made by compression or moulding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared bycompressing in a suitable machine the active ingredient in afree-flowing form such as a powder or granules, optionally mixed with abinder (e.g., povidone, gelatin, hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (e.g., sodiumstarch glycolate, cross-linked povidone, cross-linked sodiumcarboxymethyl cellulose), surface-active or dispersing agent. Mouldedtablets may be made by moulding in a suitable machine a mixture of thepowdered compound moistened with an inert liquid diluent. The tabletsmay optionally be coated or scored and may be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile. Tablets may optionally beprovided with an enteric coating, to provide release in parts of the gutother than the stomach.

Formulations suitable for topical administration (e.g., transdermal,intranasal, ocular, buccal, and sublingual) may be formulated as anointment, cream, suspension, lotion, powder, solution, paste, gel,spray, aerosol, or oil. Alternatively, a formulation may comprise apatch or a dressing such as a bandage or adhesive plaster impregnatedwith active ingredients and optionally one or more excipients ordiluents.

Formulations suitable for topical administration in the mouth includelosenges comprising the active ingredient in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as gelatin and glycerin, or sucroseand acacia; and mouthwashes comprising the active ingredient in asuitable liquid carrier.

Formulations suitable for topical administration to the eye also includeeye drops wherein the active ingredient is dissolved or suspended in asuitable carrier, especially an aqueous solvent for the activeingredient.

Formulations suitable for nasal administration, wherein the carrier is asolid, include a coarse powder having a particle size, for example, inthe range of about 20 to about 500 microns which is administered in themanner in which snuff is taken, i.e., by rapid inhalation through thenasal passage from a container of the powder held close up to the nose.Suitable formulations wherein the carrier is a liquid for administrationas, for example, nasal spray, nasal drops, or by aerosol administrationby nebuliser, include aqueous or oily solutions of the activeingredient.

Formulations suitable for topical administration via the skin includeointments, creams, and emulsions. When formulated in an ointment, theactive ingredient may optionally be employed with either a paraffinic ora water-miscible ointment base. Alternatively, the active ingredientsmay be formulated in a cream with an oil-in-water cream base. Ifdesired, the aqueous phase of the cream base may include, for example,at least about 30% w/w of a polyhydric alcohol, i.e., an alcohol havingtwo or more hydroxyl groups such as propylene glycol, butane-1,3-diol,mannitol, sorbitol, glycerol and polyethylene glycol and mixturesthereof. The topical formulations may desirably include a compound whichenhances absorption or penetration of the active ingredient through theskin or other affected areas. Examples of such dermal penetrationenhancers include dimethylsulfoxide and related analogues.

When formulated as a topical emulsion, the oily phase may optionallycomprise merely an emulsifier (otherwise known as an emulgent), or itmay comprises a mixture of at least one emulsifier with a fat or an oilor with both a fat and an oil. Preferably, a hydrophilic emulsifier isincluded together with a lipophilic emulsifier which acts as astabiliser. It is also preferred to include both an oil and a fat.Together, the emulsifier(s) with or without stabiliser(s) make up theso-called emulsifying wax, and the wax together with the oil and/or fatmake up the so-called emulsifying ointment base which forms the oilydispersed phase of the cream formulations.

Suitable emulgents and emulsion stabilisers include Tween 60, Span 80,cetostearyl alcohol, myristyl alcohol, glyceryl monostearate and sodiumlauryl sulfate. The choice of suitable oils or fats for the formulationis based on achieving the desired cosmetic properties, since thesolubility of the active compound in most oils likely to be used inpharmaceutical emulsion formulations may be very low. Thus the creamshould preferably be a non-greasy, non-staining and washable productwith suitable consistency to avoid leakage from tubes or othercontainers. Straight or branched chain, mono- or dibasic alkyl esterssuch as di-isoadipate, isocetyl stearate, propylene glycol diester ofcoconut fatty acids, isopropyl myristate, decyl oleate, isopropylpalmitate, butyl stearate, 2-ethylhexyl palmitate or a blend of branchedchain esters known as Crodamol CAP may be used, the last three beingpreferred esters. These may be used alone or in combination depending onthe properties required. Alternatively, high melting point lipids suchas white soft paraffin and/or liquid paraffin or other mineral oils canbe used.

Formulations suitable for rectal administration may be presented as asuppository with a suitable base comprising, for example, cocoa butteror a salicylate.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient, such carriers as areknown in the art to be appropriate.

Formulations suitable for parenteral administration (e.g., by injection,including cutaneous, subcutaneous, intramuscular, intravenous andintradermal), include aqueous and non-aqueous isotonic, pyrogen-free,sterile injection solutions which may contain anti-oxidants, buffers,preservatives, stabilisers, bacteriostats and solutes which render theformulation isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents, and liposomes or other microparticulatesystems which are designed to target the compound to blood components orone or more organs. Examples of suitable isotonic vehicles for use insuch formulations include Sodium Chloride Injection, Ringer's Solution,or Lactated Ringer's Injection. Typically, the concentration of theactive ingredient in the solution is from about 1 ng/ml to about 10μg/ml, for example from about 10 ng/ml to about 1 μg/ml. Theformulations may be presented in unit-dose or multi-dose sealedcontainers, for example, ampoules and vials, and may be stored in afreese-dried (lyophilised) condition requiring only the addition of thesterile liquid carrier, for example water for injections, immediatelyprior to use. Extemporaneous injection solutions and suspensions may beprepared from sterile powders, granules, and tablets. Formulations maybe in the form of liposomes or other microparticulate systems which aredesigned to target the active compound to blood components or one ormore organs.

Dosage

It will be appreciated that appropriate dosages of the active compounds,and compositions comprising the active compounds, can vary from patientto patient. Determining the optimal dosage will generally involve thebalancing of the level of therapeutic benefit against any risk ordeleterious side effects of the treatments of the present invention. Theselected dosage level will depend on a variety of factors including, butnot limited to, the activity of the particular compound, the route ofadministration, the time of administration, the rate of excretion of thecompound, the duration of the treatment, other drugs, compounds, and/ormaterials used in combination, and the age, sex, weight, condition,general health, and prior medical history of the patient. The amount ofcompound and route of administration will ultimately be at thediscretion of the physician, although generally the dosage will be toachieve local concentrations at the site of action which achieve thedesired effect.

Administration in vivo can be effected in one dose, continuously orintermittently throughout the course of treatment. Methods ofdetermining the most effective means and dosage of administration arewell known to those of skill in the art and will vary with theformulation used for therapy, the purpose of the therapy, the targetcell being treated, and the subject being treated. Single or multipleadministrations can be carried out with the dose level and pattern beingselected by the treating physician.

In general, a suitable dose of the active compound is in the range ofabout 0.1 to about 250 mg per kilogram body weight of the subject perday. Where the active ingredient is a salt, an ester, prodrug, or thelike, the amount administered is calculated on the basis the parentcompound and so the actual weight to be used is increasedproportionately.

Prodrugs

Compounds of the present invention may be prodrugs for potentantiproliferative agents. Compounds which exhibit low or moderateintrinsic activity may act as prodrugs, and be metabolically activated(e.g., in vivo) to generate more potent compounds. This is especiallyuseful in cancer therapy where metabolic activation can be achieved byan enzyme that is expressed in tumours.

For example, the cytochrome P450 enzyme CYP1B1 has been shown to bespecifically expressed in tumour cells, but is not found in thecorresponding normal tissues. This enzyme is found to be expressed in avariety of tumours, such as brain, breast, colon, stomach, ovarian andprostate cancers (see, e.g., Murray et al, 1997; Melvin et al., 1997).Prodrugs, acting as a substrate, may be metabolised by CYP1B1 through anaromatic hydroxylation reaction to generate a potent anticancer agent.

For example, as illustrated below, a prodrug, with low intrinsicactivity (e.g., IC50 of 0.69 μM in breast cancer MCF-7 cells)(E)-1-(4-methoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one, isconverted to the hydroxylated metabolite,(E)-1-(3-Hydroxy-4-methoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one,which has substantially potency (e.g., IC50 of 0.00065 μM in the samecell line).

Thus, those compounds of the present invention where R^(A3) is —H may beprodrugs, to be activated by CYP1B1 enzyme, to yield the correspondingdrug where R^(A3) is —OH.

In such cases, the prodrug is useful as an antiproliferative agent withlow intrinsic toxicity, for treatment of proliferative conditionscharacterised by cells which express the CYP1B1 enzyme.

Additionally, the prodrug is useful as a selective antiproliferativeagent with low intrinsic toxicity, for treatment of proliferativeconditions characterised by cells which express the CYP1B1 enzyme, wherethe corresponding normal cells do not express the CYP1B1 enzyme.

Furthermore, prodrugs with low intrinsic cytotoxicity, which are onlyactivated upon entering cells (e.g., tumour cells) containing the CYP1B1enzyme, are not only useful for treating cancer, but also as aprophylactic, in cancer prevention (i.e., as a cancer preventativeagent).

A method for detecting and/or demonstrating the conversion of acandidate prodrug to the corresponding drug is described next: Amicrosomal preparation of human tumour tissue expressing the CYP1B1enzyme is prepared essentially as described by the method of Barrie etal., 1989. The experiment is carried out at 37° C., under yellow light.An array of 1.5 ml centrifuge tubes are set up in a water bath shakerunder aerobic conditions. To each tube is then added 500 μl of pH 7.6buffer (0.1 M NaK₂PO₄), followed by NADPH (5 μl of a 25 mM stocksolution). The microsomal preparation (80 μl) is then added and thetubes pre-incubated for 5 min at 37° C. The prodrug is then added (10 μlof a 5 mM stock solution) and the preparation incubated for 1 h at 37°C. After 1 h the tubes are transferred to an ice/water cooling bath (0°C.). The tubes are then centrifuged at 15,000 rpm for 30 min. A sampleof the supernatant (100 μl) is then taken and analysed by HPLC. HPLCconditions: Spherisorb C18 (25 cm×4.6 mm id), used without guard column.Flow rate 1 ml/min. Eluent 75% 0.1 M KH₂PO₄ and 25% acetonitrile. Thehydroxylated drug is detected by HPLC, and confirmed by comparison withthe authentic hydroxylated synthetic compound.

Diagnosis and Assays

In many cases, hydroxylated compounds, where R^(A3) is —OH, exhibit muchgreater fluorescence than the corresponding non-hydroxylated compound,where R^(A3) is —H. This property may be exploited in diagnosis, forexample, of cancer, by detecting and/or measuring the formation of thehydroxylated metabolite via tumour cells expressing the CYP1B1 enzyme.

Thus, one aspect of the present invention pertains to a method ofdiagnosis of a subject for the presence of cells (e.g., tumour cells)expressing the CYP1B1 enzyme, comprising:

-   -   (a) administering to the patient a non-hydroxylated prodrug as        described herein, wherein R^(A3) is —H;    -   (b) determining the amount of the corresponding hydroxylated        metabolite, wherein R^(A3) is —OH which is subsequently        produced; and,    -   (c) correlating the amount with the presence or absence of the        cells in the patient.

Another aspect of the present invention pertains to active compounds,wherein R^(A3) is —H, for use in a method of diagnosis of the human oranimal body. In one embodiment, the diagnosis is for the presence ofcells (e.g., tumour cells) expressing the CYP1B1 enzyme.

Another aspect of the present invention pertains to use of activecompounds, wherein R^(A3) is —H, for the manufacture of a composition,for example, for the diagnosis of the presence of cells (e.g., tumourcells) expressing the CYP1B1 enzyme, a proliferative condition, aninflammatory condition, etc., as discussed above.

Kits

One aspect of the invention pertains to a kit comprising (a) the activeingredient, preferably provided in a suitable container and/or withsuitable packaging; and (b) instructions for use, for example, writteninstructions on how to administer the active compound, how to perform adiagnosis using the active compound, etc.

The written instructions may also include a list of indications forwhich the active ingredient is a suitable treatment.

EXAMPLES

The following are examples are provided solely to illustrate the presentinvention and are not intended to limit the scope of the invention, asdescribed herein.

Analytical Methods

The ¹H- and ¹³C-NMR spectra were recorded on a 250 MHz super-conductingBruker AC250 Spectrometer. Infrared spectra were recorded in potassiumbromide on a Shimadzu FTIR-8300 Spectrophotometer. The mass spectra wererecorded on a VG 70 SEQ Spectrometer. Melting points were determined onan Electrothermal melting point apparatus and were uncorrected. Thinlayer chromatography was performed on silica gel sheets (Merck TLCAluminium sheet-Silica Gel 60F) and was monitored with UV light. Columnchromatography was performed using Silica gel 60 (220–440 mesh).

Example 1 (E)-1-(4-ethoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one(DMU-174)

To a stirred solution of 4-ethoxybenzaldehyde (1.0 g, 6.6 mmol) and3,5-dimethoxyacetophenone (1.19 g, 6.6 mmol) in methanol (10 ml) wasadded 50% w/v of aqueous sodium hydroxide (5.3 ml, 0.06 mol). Thereaction mixture was stirred for 2 h at room temperature and thenextracted with ethyl acetate (3×20 ml). The combined organic layers weredried over magnesium sulphate and reduced in vacuo and the product wasrecrystallised from ethanol to give 1.12 g (74%) of yellow crystals.¹H-NMR (CDCl₃) δ 7.8 (d, 1H), 7.6 (d, 2H), 7.3 (d, 1H), 7.15 (d, 2H),6.9 (d, 2H), 6.65 (t, 1H), 4.0 (q, 2H), 3.8 (s, 6H), 1.4 (t, 3H).¹³C-NMR (CDCl₃) δ 190.1, 161.1, 160.8, 140.6, 127.4, 63.6. Mass Spectrumm/e (M+1) 313.

Example 2 (E)-1-(4-propoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one(DMU-175)

To a stirred solution of 4-propoxybenzaldehyde (0.5 g, 3.1 mmol) and3,5-dimethoxyacetophenone (0.55 g, 3.1 mmol) in methanol (10 ml) wasadded 50% w/v of aqueous sodium hydroxide (2.4 ml, 0.031 mol). Thereaction mixture was stirred for 2 h at room temperature and thenextracted with ethyl acetate (3×20 ml), the combined organic layers weredried over magnesium sulphate and reduced in vacuo. The product wasrecrystallised from ethanol to give 0.978 g (93%) of off-white crystals.¹H-NMR (CDC₃) δ 7.8 (d, 1H), 7.6(d, 2H), 7.3 (d, 1H), 7.15 (d, 2H), 6.9(d, 2H), 6.65 (t, 1H), 4.0 (t, 2H), 3.9 (s, 6H), 1.85 (m, 2H), 1.05 (t,3H). ¹³C-NMR (CDCl₃) δ 190.2, 161.3, 160.8, 144.9, 140.6, 130.2, 127.4,119.6, 114.9, 106.3, 104.8, 69.7, 55.6, 22.5, 10.4. Mass Spectrum m/e(M+1)327.

Example 3 (E)-1-(4-butoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one(DMU-176)

To a stirred solution of 4-butoxybenzaldehyde (0.5 g, 2.8 mmol) and3,5-dimethoxyacetophenone (0.5 g, 2.8 mmol) in methanol (10 ml) wasadded 50% w/v of aqueous sodium hydroxide (2.24 ml, 0.028 mol). Thereaction mixture was stirred for 2 h at room temperature and thenextracted with ethyl acetate (3×20 ml), the combined organic layers weredried over magnesium sulphate and reduced in vacuo. The product wasrecrystallised from ethanol to give 0.925 g (97%) of off-white crystals.¹H-NMR (CDCl₃) δ 7.8 (d, 1H), 7.6 (d, 2H), 7.35 (d, 1H), 7.1 (d, 2H),6.9 (d, 2H), 6.7 (t, 1H), 4.0 (t, 2H), 3.85 (s, 6H), 1.75 (m, 2H), 1.5(m, 2H), 1.0 (t, 3H). ¹³C-NMR (CDCl₃) δ 191.1, 161.4, 160.8, 144.9,140.8, 130.2, 127.3, 119.6, 114.9, 106.3, 104.8, 67.9, 55.6, 31.2, 19.2,13.8. Mass Spectrum m/e (M+1) 341.

Example 4(E)-1-(3-hydroxy-4-ethoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one(DMU-184)

Anhydrous potassium carbonate (2.25 g, 16.3 mmol) was added to3,4-dihydroxybenzaldehyde (1.5 g, 10.8 mmol), in butanone (30 ml), atroom temperature. The reaction was stirred for 10 min after which ethyliodide (0.872 ml, 10.8 mmol) was added dropwise and the reaction wasrefluxed for 3 h. The reaction mixture was diluted with water and theproduct was extracted with dichloromethane (3×25 ml), the combinedorganic layers were dried over magnesium sulphate and reduced in vacuo.Purification by column chromatography (SiO₂, petroleum:ether (40:60,v/v) with an increasing gradient of ethyl acetate) afforded 0.78 g (43%)of 3-hydroxy-4-ethoxybenzaldehyde as a white solid.

A mixture of 3-hydroxy-4-ethoxybenzaldehyde (0.413 g, 2.5 mmol),3,5-dimethoxyacetophenone (0.448 mg, 2.5 mmol) and 50% w/v of aqueoussodium hydroxide (3.98 ml, 0.049 mol) in methanol (5 ml) at roomtemperature for 18 h. The yellow solid that precipitated was filteredand sequentially washed with cold methanol and ether and finally driedin a vacuum dessicator. Purification by column chromatography (SiO₂,petroleum:ether (40:60 v/v) with an increasing gradient of ethylacetate) yielded 0.29 g (36%) of a yellow solid. ¹H-NMR (CDCl₃) δ 7.7(d, 1H), 7.2 (m, 2H), 7.1 (d, 3H), 6.8 (d, 1H), 6.5 (t, 1H), 5.7 (s,1H), 4.1 (q, 2H), 3.9 (s, 6H), 1.4 (t, 3H). ¹³C-NMR (CDCl₃) δ 190.1,160.9, 148.2, 146.1, 144.9, 140.6, 128.5, 122.7, 120.3, 113.1, 111.4,106.3, 105.0, 64.7, 55.7, 14.8. Mass Spectrum m/e (M+1) 329. Anal.Calcd. for C₁₉H₂₀O₅.0.5H₂O: C, 69.5; H, 6.14. Found C, 69.62; H, 6.16.

Example 5(E)-1-(3-hydroxy-4-propoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one(DMU-185)

Anhydrous potassium carbonate (1.5 g, 10.8 mmol) and sodium iodide(0.108 g, 0.72 mmol) were added to 3,4-dihydroxybenzaldehyde (1.0 g, 7.2mmol), in butanone (50 ml), at room temperature. The reaction wasstirred for 10 min after which propyl bromide (0.657 ml, 7.2 mmol) wasadded dropwise and the reaction was refluxed for 3 h. The reactionmixture was diluted with water and the product was extracted withdichloromethane (3×25 ml), the combined organic layers were dried overmagnesium sulphate and reduced in vacuo. Purification by columnchromatography (SiO₂, petroleum:ether (40:60 v/v) with an increasinggradient of ethyl acetate) afforded 0.248 g (22%) of3-hydroxy-4-propoxybenzaldehyde as an off-white solid.

A mixture of 3-hydroxy-4-propoxybenzaldehyde (0.4 g, 2.1 mmol),3,5-dimethoxyacetophenone (0.371 mg, 2.1 mmol) and 50% w/v of aqueoussodium hydroxide (3.3 ml, 0.206 mol) in methanol (10 ml) at roomtemperature for 18 h. The yellow solid that precipitated was filteredand sequentially washed with cold methanol and ether and finally driedin a vacuum dessicator. Purification was achieved using columnchromatography (SiO₂, petroleum:ether (40:60 v/v) with an increasinggradient of ethyl acetate) which gave 0.95 g (19%) of a yellow solid.¹H-NMR (CDCl₃) δ 7.7 (d, 1H), 7.3 (m, 2H), 7.1 (d, 3H), 6.8 (d, 1H), 6.6(d, 1H), 5.7 (s, 1H), 4.1 (t, 2H), 3.9 (s, 6H), 1.9 (m, 2H), 1.0 (t,3H). ¹³C-NMR (CDCl₃) δ 160.9, 146.0, 144.9, 128.4, 122.6, 120.2, 112.9,106.2, 104.9, 70.6, 55.6, 22.4, 10.4. Mass Spectrum m/e (M+1) 343. Anal.Calcd. for C₂₀H₂₂O₅: C, 70.16; H, 6.48. Found C, 70.08; H, 6.38.

Example 6(E)-1-(3-hydroxy-4-butoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one(DMU-186)

Anhydrous potassium carbonate (1.5 g, 5.4 mmol) and sodium iodide (0.108g, 0.36 mmol) were added to 3,4-dihydroxybenzaldehyde (0.5 g, 3.6 mmol),In butanone (25 ml), at room temperature. The reaction was stirred for10 min after which butyl bromide (0.777 ml, 3.6 mmol) was added dropwiseand the reaction was refluxed for 3 h. The reaction mixture was dilutedwith water and the product was extracted with dichloromethane (3×25 ml),the combined organic layers were dried over magnesium sulphate andreduced in vacuo. Purification by column chromatography (SiO₂,petroleum:ether (40:60 v/v) with an increasing gradient of ethylacetate) afforded 0.5 g (71%) of 3-hydroxy-4-butoxybenzaldehyde as anoff-white solid.

A mixture of 3-hydroxy-4-butoxybenzaldehyde (0.4 g, 2.1 mmol),3,5-dimethoxyacetophenone (0.371.9, 2.1 mmol) and 50% w/v of aqueoussodium hydroxide (3.3 ml, 0.206 mol) in methanol (10 ml) at roomtemperature for 18 h. The yellow solid that precipitated was filteredand sequentially washed with cold methanol and ether and finally driedin a vacuum dessicator. Purification was achieved using columnchromatography (SiO₂, petroleum:ether (40:60 v/v) with an increasinggradient of ethyl acetate) which gave 0.36 g (50%) of a yellow solid:¹H-NMR (CDCl₃) δ 7.7 (d, 1H), 7.3 (m, 2H), 7.1 (d, 3H), 6.8 (d, 1H), 6.7(t, 1H), 5.7 (s, 1H), 4.1 (t, 2H), 3.9 (s, 6H), 1.8 (m, 2H), 1.5 (m,2H), 1.0 (t, 3H). ¹³C-NMR (CDCl₃) δ 190.0, 160.9, 148.3, 146.0, 144.9,140.5, 128.4, 122.6, 120.2, 112.9, 111.4, 106.2, 104.9, 68.8, 55.6,31.1, 19.1. 13.7. Mass Spectrum m/e (M+1) 357. Anal. Calcd. forC₂₁H₂₄O₅: C, 70.77, H, 6.79. Found C, 70.84; H, 6.81.

Example 7(E)-1-(4-ethoxyphenyl)-3-(3,4,5-trimethoxyphenyl)prop-1-en-3-one(DMU-190)

To a stirred solution of 4-ethoxybenzaldehyde (1.32 ml, 9.5 mmol) and3,4,5-trimethoxyacetophenone (2.0 g, 9.5 mmol) in methanol (30 ml) wasadded 50% w/v of aqueous sodium hydroxide (7.6 ml, 0.095 mol). Thereaction was stirred for 24 h at room temperature. A precipitate wasisolated by filtration, washed with pet:ether and subsequentlyrecrystallised from ethanol. 1.8 g (55%) of a pale yellow powder wasisolated. ¹H-NMR (CDCl₃) δ 7.8 (d, 1H), 7.6 (d, 2H), 7.3 (d, 1H), 6.9(d, 2H), 4.1 (q, 2H), 3.9 (s×2, 9H), 1.4 (t, 3H). ¹³C-NMR (CDCl₃) δ189.2, 161.1, 153.1, 142.3, 133.8, 127.4, 63.4. Mass Spectrum m/e (M+1)343.

Example 8(E)-1-(4-propoxyphenyl)-3-(3,4,5-trimethoxyphenyl)prop-1-en-3-one(DMU-191)

To a stirred solution of 4-propoxybenzaldehyde (1.5 ml, 9.5 mmol) and3,4,5-trimethoxyacetophenone (2.0 g, 9.5 mmol) in methanol (30 ml) wasadded 50% w/v of aqueous sodium hydroxide (7.6 ml, 0.095 mol). Thereaction was stirred for 24 h at room temperature. A precipitate wasisolated by filtration, washed with pet:ether and subsequentlyrecrystallised from ethanol to give 0.9 g (27%) of a pale yellow powder.¹H-NMR (CDCl₃) δ 7.8 (d, 1H), 7.6 (d, 2H), 7.3 (d, 1H), 7.2 (m, 1H), 6.8(d, 2H), 3.95 (m, 2H), 3.9 (s×2, 9H), 1.8 (t, 2H), 1.0 (t, 3H).

¹³C-NMR (CDCl₃) δ 189.3, 161.3, 156.9, 153.1, 144.7, 142.4, 133.8,130.1, 127.4, 119.3, 114.9, 106.1, 69.7, 60.9, 56.3, 22.5, 10.4. MassSpectrum m/e (M+1) 357.

Example 9(E)-1-(4-butoxyphenyl)-3-(3,4,5-trimethoxyphenyl)prop-1-en-3-one(DMU-192)

To a stirred solution of 4-butoxybenzaldehyde (1.64 ml, 9.5 mmol) and3,4,5-trimethoxyacetophenone (2.0 g, 9.5 mmol) in methanol (30 ml) wasadded 50% w/v of aqueous sodium hydroxide (7.6 ml, 0.095 mol). Thereaction was stirred for 24 h at room temperature. A precipitate wasisolated by filtration, washed with pet:ether and subsequentlyrecrystallised from ethanol to give 3.0 g (85%) of a pale yellow powder.¹H-NMR (CDCl₃) δ 7.8 (d, 1H), 7.6 (d, 2H), 7.4 (d, 1H), 7.25 (d, 2H),7.0 (d, 2H), 4.05 (t, 2H), 4.0 (s×2, 9H), 1.8 (m, 2H), 1.5 (m, 2H), 1.0(t, 3H). Mass Spectrum m/e (M+1) 370.

Example 10(E)-1-(3-hydroxy4-ethoxyphenyl)-3-(3,4,5-trimethoxyphenyl)prop-1-en-3-one(DMU-187)

Anhydrous potassium carbonate (2.25 g, 16.3 mmol) was added to3,4-dihydroxybenzaldehyde (1.5 g, 10.8 mmol), in butanone (30 ml), atroom temperature. The reaction was stirred for 10 min after which ethyliodide (0.872 ml, 10.8 mmol) was added dropwise and the reaction wasrefluxed for 3 h. The reaction mixture was diluted with water and theproduct was extracted with dichloromethane (3×25 ml), the combinedorganic layers were dried over magnesium sulphate and reduced in vacuo.Purification by column chromatography (SiO₂, petroleum:ether (40:60)with an increasing gradient of ethyl acetate) afforded 0.78 g (43%) of3-hydroxy4-ethoxybenzaldehyde as a white solid.

A mixture of 3-hydroxy-4-ethoxybenzaldehyde (0.350 g, 2.5 mmol),3,4,5-trimethoxyacetophenone (0.443 mg, 2.5 mmol) and 50% w/v of aqueoussodium hydroxide (3.37 ml, 0.042 mol) in methanol (5 ml) at roomtemperature for 18 h. An orange solid was isolated by filtration,dissolved in ethyl acetate which was subsequently dried over MgSO₄ andreduced in vacuo. Purification by column chromatography (SiO₂,petroleum:ether (40:60 v/v) with an increasing gradient of ethylacetate) yielded 0.126 g (16.7%) of a yellow solid. ¹H-NMR (CDCl₃) δ 7.7(d, 1H), 7.3 (m, 4H), 7.1 (dd, 1H), 6.9 (d, 1H), 5.75 (s, 1H), 4.2 (q,2H), 3.95 (s×2, 9H), 1.5 (t, 3H). ¹³C-NMR (CDCl₃) δ 190.0, 153.1, 150.0,146.0, 133.7, 128.4. Mass Spectrum m/e (M+1) 359. Anal. Calcd. forC₂₀H₂₂O₆: C, 67.03; H, 6.19. Found C, 66.74; H, 6.33.

Example 11(E)-1-(3-hydroxy4-propoxyphenyl)-3-(3,4,5-trimethoxyphenyl)prop-1-en-3-one(DMU-188)

Anhydrous potassium carbonate (5.9 g, 0.043 mmol) and sodium iodide(0.434 g, 4.3 mmol) were added to 3,4-dihydroxybenzaldehyde (4.0 g, 28.9mmol), in butanone (100 ml), at room temperature. The reaction wasstirred for 10 min after which propyl bromide (2.628 ml, 28.9 mmol) wasadded dropwise and the reaction was refluxed for 3 h. The reactionmixture was diluted with water and the product was extracted withdichloromethane (3×50 ml), the combined organic layers were dried overmagnesium sulphate and reduced in vacuo. Purification by columnchromatography (SiO₂, petroleum:ether (40:60 v/v) with an increasinggradient of ethyl acetate) afforded 1.21 g (23.3%) of3-hydroxy-4-propoxybenzaldehyde as an off-white solid.

A mixture of 3-hydroxy-4-propoxybenzaldehyde (0.8 g, 4.4 mmol),3,4,5-trimethoxyacetophenone (0.934 mg, 4.4 mmol) and 50% w/v of aqueoussodium hydroxide (7.0 ml, 0.089 mol) in methanol (10 ml) at roomtemperature for 18 h. The orange solid isolated by filtration was washedsequentially with cold methanol and ether and finally dried in a vacuumdessicator. Purification was achieved using column chromatography (SiO₂,petroleum:ether (40:60 v/v) with an increasing gradient of ethylacetate) which gave 0.995 g (60%) of a yellow solid. ¹H-NMR (CDCl₃) δ7.7 (d, 1H), 7.2 (m, 4H), 7.1 (dd, 3H), 6.9 (d, 1H), 5.7 (s, 1H), 4.1(t, 2H), 3.9 (s×2, 9H), 1.9 (m, 2H), 1.1 (t, 3H). ¹³C-NMR (CDCl₃) δ189.1, 153.1, 148.3, 146.0, 144.7, 142.4, 133.7, 128.4, 122.9, 119.8,112.7, 111.4, 106.1, 70.6, 60.9, 56.4, 22.4, 10.4. Mass Spectrum m/e(M+1) 373. Anal. Calcd. for C₂₁H₂₄O₆: C, 67.73; H, 6.5. Found C, 68.01;H, 6.72.

Example 12(E)-1-(3-hydroxy-4-butoxyphenyl)-3-(3,4,5-trimethoxyphenyl)prop-1-en-3-one(DMU-189)

Anhydrous potassium carbonate (5.9 g, 0.0434 mmol) and sodium iodide(0.434 g, 2.9 mmol) were added to 3,4-dihydroxybenzaldehyde (4.0 g, 28.9mmol), in butanone (100 ml), at room temperature. The reaction wasstirred for 10 min after which butyl bromide (3.11 ml, 28.9 mmol) wasadded dropwise and the reaction was refluxed for 3 h. The reactionmixture was diluted with water and the product was extracted withdichloromethane (3×50 ml), the combined organic layers were dried overmagnesium sulphate and reduced in vacuo. Purification by columnchromatography (SiO₂, petroleum:ether (40:60 v/v) with an increasinggradient of ethyl acetate) afforded 2.25 g (58%) of3-hydroxy-4-butoxybenzaldehyde as an off-white solid.

A mixture of 3-hydroxy-4-butoxybenzaldehyde (1.0 g, 5.2 mmol),3,4,5-trimethoxyacetophenone (1.08 g, 5.2 mmol) and 50% w/v of aqueoussodium hydroxide (8.25 ml, 0.103 mol) in methanol (10 ml) at roomtemperature for 18 h. An orange solid was isolated by filtration and wassequentially washed with cold methanol and ether and finally dried in avacuum dessicator. Purification was achieved using column chromatography(SiO₂, petroleum:ether (40:60 v/v) with an increasing gradient of ethylacetate) which gave 0.224 g (11.3%) of a yellow solid: ¹H-NMR (CDCl₃) δ7.7 (d, 1H), 7.3 (m, 4H), 7.1 (dd, 1H), 6.9 (d, 1H), 5.7 (t, 1H), 5.7(s, 1H), 4.1 (t, 2H), 4.0 (s×2, 9H), 1.8 (m, 2H), 1.5 (m, 2H), 1.0 (t,3H). ¹³C-NMR (CDCl₃) δ 189.1, 157.6, 153.2, 146.1, 144.7, 133.8, 122.9,119.9, 112.7, 111.4, 106.1, 68.9, 60.9, 56.4, 31.2, 19.2, 15.01, 13.8.Mass Spectrum m/e (M+1) 387. Anal. Calcd. for C₂₂H₂₆O₆: C, 68.38; H,6.78. Found C, 68.57; H, 7.01.

Example 13(E)-1-(4-isopropoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one(DMU-401)

To a stirred solution of 4-isopropoxybenzaldehyde (1.0 g, 6.09 mmol) and3,5-dimethoxyacetophenone (1.1 g, 6.09 mmol) in methanol (30 ml) wasadded a 50% w/v solution of aqueous sodium hydroxide (11 ml, 0.138 mol).The reaction mixture was stirred for 24 h at room temperature; noprecipitate was observed. The product was extracted with ethyl acetate(3×50 ml), the combined organic layers were dried over anhydrousmagnesium sulphate and reduced in vacuo. Purification by columnchromatography (SiO₂, petroleum:ether (40:60 v/v) with an increasinggradient of ethyl acetate) afforded a yellow oil which was trituratedwith hexane to give a pale yellow solid (1.14 g, 57%). ¹H-NMR (CDCl₃) δ7.7 (d, 1H), 7.5 (m, 2H), 7.3 (d, 1H), 7.1 (d, 2H), 6.9 (m, 2H), 6.5 (t,1H) 4.6 (m, 1H), 3.85 (s, 6H), 1.35 (d, 6H). ¹³C-NMR (CDCl₃) δ 190.2,160.8, 140.6, 129.0, 127.2, 117.2, 114.6, 107.5, 56.7. Mass Spectrum m/e(M+1) 327. Anal. Calcd. For C₂₀H₂₂O₄.0.5 H₂O: C, 73.59; H, 6.80. FoundC, 71.62; H, 6.91.

Example 14 (E)-1-(4-propoxyphenyl)-3-(3-methoxyphenyl)prop-1-en-3-one(DMU-417)

To a stirred solution of 4-propoxybenzaldehyde (1 g, 6.09 mmol) and3-methoxyacetophenone (0.91 g, 6.09 mmol) in methanol (30 ml) was added50% w/v solution of aqueous sodium hydroxide (9.7 ml, 0.122 mol). Thereaction mixture was stirred at room temperature for 24 h. A precipitatewas isolated by filtration, washed with cold methanol and subsequentlyrecrystallised from methanol to afford 0.38 g (21%) of pale yellowcrystals. ¹H-NMR δ (CDCl₃), 1.6 (t, 3H), 1.85 (m, 2H), 3.9 (s, 2H), 3.95(t, 2H), 6.9 (d, 2H), 7.1 (1H), 7.38 (m, 2H), 7.55 (m, 4H), 7.8 (d, 1H).¹³C-NMR (CDCl₃) δ 190.3, 161.3, 159.9, 139.9, 127.4, 69.6, 22.5. MassSpectrum m/e (M+1) 297. Anal. Calcd. for C₁₉H₂₀O₃: C, 77.00; H, 6.80.Found C, 76.86; H, 6.94.

Example 15(E)-1-(4-propoxyphenyl)-3-(2,4-dimethoxyphenyl)prop-1-en-3-one (DMU-408)

To a stirred solution of 4-propoxybenzaldehyde (1.0 g, 6.1 mmol) and2,′4′-dimethoxyacetophenone (1.1 g, 6.1 mmol) in methanol (30 ml) wasadded 50% w/v solution of aqueous sodium hydroxide (9.7 ml, 0.122 mol).The reaction mixture was stirred at room temperature for 2 h. Aprecipitate was isolated by filtration and subsequently recrystallisedfrom methanol to afford 0.31 g (16%) of pale yellow crystals. ¹H-NMR(CDCl₃) δ 1.0 (t, 3H), 1.7 (m, 2H), 3.85 (s, 3H), 3.9 (s, 3H), 3.9 (t,2H), 6.5 (dd, 2H), 6.85 (d, 2H), 7.3 (d, 1H), 7.6 (d, 2H), 7.5 (d, 2H).¹³C-NMR (CDCl₃) δ 190.6, 163.9, 160.7, 160.2, 127.9, 122.5, 69.6, 22.5.Mass Spectrum m/e (M+1) 327. Anal. Calcd. for C₂₀H₂₂O₄: C, 73.62; H,6.75. Found C, 73.48; H, 6.81.

Example 16(E)-1-(4-propoxyphenyl)-3-(2,3,4-trimethoxyphenyl)prop-1-en-3-one(DMU-420)

To a stirred solution of 4-propoxybenzaldehyde (0.781 g, 4.8 mmol) and2,3,4-trimethoxyacetophenone (1.0 g, 4.8 mmol) in methanol (30 ml) wasadded a 50% w/v solution of aqueous NaOH (7.60 ml, 0.096 mol). Thereaction mixture was stirred at room temperature for 24 h. A pale yellowprecipitate was isolated and dried under vacuum over night. 1.693 g(86%) of a pale yellow powder was isolated. ¹H-NMR (CDCl₃) δ 7.60 (d,1H), 7.5 (m, 1H), 4.4 (d, 1H), 7.3 (d, 1H), 6.8 (m, 1H), 6.7 (d, 1H),3.9 (t, 2H), 3.9 (s, 3H), 3.9 (s, 3H), 3.9 (s, 3H), 1.75 (m, 2H), 1.0(t, 3H). ¹³C-NMR (CDCl₃) δ 191.0. Mass Spectrum m/e (M+1) 357. Anal.Calcd. for C₂₁H₂₄O₅.H₂O: C, 67.44; H, 7.01. Found: C, 67.49; H, 6.93.

Example 17(E)-1-(4-propoxyphenyl)-3-(2,5-dimethoxyphenyl)prop-1-en-3-one (DMU-424)

To a stirred solution of 4-propoxybenzaldehyde (1 g, 6.1 mmol) and2′,5′-dimethoxyacetophenone (1.1 g, 6.1 mmol) in methanol (15 ml) wasadded 50% w/v solution of aqueous sodium hydroxide (9.7 ml, 0.122 mol).The reaction mixture was stirred at room temperature for 18 h. Aprecipitate was isolated by filtration and subsequently recrystallisedfrom ethanol to afford 0.97 g (49%) of pale yellow crystals. ¹H-NMR(CDCl₃) δ 1.0 (t, 3H), 1.8 (m, 2H), 3.7 (s, 3H), 3.8 (s, 3H), 3.9 (t,2H), 6.9 (m, 3H), 6.9 (d, 1H) 7.15 (d, 1H), 7.25 (d, 1H), 7.5 (m, 3H),¹³C-NMR (CDCl₃) δ 192.59, 160.86,161.11, 153.59, 152.34, 130.07, 127.56,22.47. Mass Spectrum m/e (M+1) 327. Anal. Calcd. for C₂₀H₂O₄: C, 73.62;H, 6.75. Found C, 73.55; H, 6.88.

Example 18(E)-1-(4-propoxyphenyl)-2-methyl-3-(3,5-dimethoxyphenyl)prop-1-en-3-one(DMU-432)

3,5-Dimethoxybenzaldehyde (4.0 g, 0.024 mol), in dry tetrahydrofuran(100 ml), was added over 15 min to ethylmagnesium bromide (28.9 ml (1.0M solution in tetrahydrofuran) 0.029 mmol) in dry tetrahydrofuran (50ml) at 0° C., under nitrogen. After refluxing the mixture for 18 h agrey solution was obtained. The reaction was then quenched by adding iceand 1 M hydrochloric acid (100 ml) dropwise and the aqueous phase wasextracted with ether (3×100 ml), the combined organic layers were driedover anhydrous magnesium sulphate and reduced in vacuo. Purification bycolumn chromatography (SiO₂, petroleum:ether (40:60 v/v) with anincreasing gradient of ethyl acetate (0–20%)) gave 3.53 g (74.8%) of thealcohol as a yellow oil.

To a stirred solution of dimethylsulfoxide (2.86 ml, 0.040 mol) in drydichloromethane (30 ml) at −78° C. was added, over 15 min, oxalylchloride (1.75 ml, 0.020 mol) under nitrogen. The solution was stirredfor 15 min at −78° C. until the evolution of gas stopped, then asolution of the alcohol (3.5 g, 0.018 mol) in dichloromethane (30 ml)was added over 15 min. The mixture was stirred at −78° C. for a further30 min before triethylamine (12.47 ml, 0.09 mol) was added over 10 min,this was stirred for a further 5 min at −78° C. and then allowed to warmup to room temperature and left for 2 h. The mixture was then dilutedwith dichloromethane (30 ml) and the organic layer was sequentiallywashed with 1 M hydrochloric acid (2×50 ml), water (2×50 ml), dried overmagnesium sulphate and reduced in vacuo. Purification by columnchromatography (SiO₂, petroleum:ether (40:60 v/v) with an increasinggradient of ethyl acetate) gave 2.85 g (81%) of the ketone as a yellowsolid.

A mixture of the ketone (1.55 g, 7.9 mmol), 4-propoxybenzaldehyde (1.24ml, 7.8 mmol), piperidine (1.76 ml, 17.8 mmol) and glacial acetic acid(0.883 ml, 15 mmol) in dry ethanol (100 ml) were heated under reflux andwater was removed from the reaction by soxhlet extraction over 4Amolecular sieves for 50 h. The solvent was removed in vacuo and theresidue was purified by column chromatography (SiO₂, petroleum:ether(40:60 v/v) with an increasing gradient of ethyl acetate) to give 0.20 g(7%) of a yellow oil. ¹H-NMR (CDCl₃) δ 7.3 (d, 2H), 7.1 (s, 1H), 6.8 (d,2H), 6.7 (d, 2H), 6.5 (t, 1H), 3.8 (t, 2H), 3.7 (s, 6H), 2.2 (t, 3H),1.7 (q, 2H), 0.9 (t, 3H). ¹³C-NMR (CDCl₃) δ 198.9, 160.5, 159.6, 142.9,140.9, 134.4, 131.6, 128.0, 126.9, 114.5, 107.2, 103.5, 69.5, 55.4,42.9, 38.7, 31.6, 22.5, 19.7, 14.3, 10.5.

(Comparative: Tertiary Alkyl) Example 1(E)-1-(4-tertbutoxyphenyl)-3-(3,5-dimethoxyphenyl)prop-1-en-3-one(DMU-402)

To a stirred solution of 3,5-dimethoxyacetophenone (1.0 g, 5.55 mmol)and 4-t-butoxybenzaldehyde (0.99 g, 5.55 mmol) in methanol (30 ml) wasadded a 50% w/v solution of aqueous sodium hydroxide (8.8 ml, 0.11 mol).The reaction mixture was stirred for 4 h at room temperature; noprecipitate was observed. The product was extracted with ethyl acetate(3×50 ml), the combined organic layers were dried over anhydrousmagnesium sulphate and reduced in vacuo. Purification by columnchromatography (SiO₂, petroleum:ether (40:60 v/v) with an increasinggradient of ethyl acetate) afforded 1.19 g (63%) of a yellow oil. ¹H-NMR(CDCl₃) δ 7.7 (d, 1H), 7.5 (m, 2H), 7.36 (d, 1H), 7.1 (d, 2H), 7.0 (m,2H), 6.6 (t, 1H), 3.8 (s, 6H), 1.4 (s, 9H). ¹³C-NMR (CDCl₃) δ 190.0,160.9, 158.2, 140.4, 130.7, 129.5, 128.2, 124.9, 122.3, 119.4, 107.5,104.9, 79.4, 56.7, 54.4, 29.9, 27.9. Mass Spectrum m/e (M+1) 341. Anal.Calcd. for C₂₁H₂₄O₄.0.5 H₂O: C, 74.09; H, 7.10. Found C, 72.18; H, 7.21.

Biological Activity

TCDD-Induced MCF-7 Cell Line Versus MCF-7 Cell Line Cytotoxicity Assay

CYP1A1 and CYP1B1 enzyme activity is induced by TCDD(tetrachlorodibenzodioxin (Dioxin)) in breast tumour MCF-7 cells (see,e.g., Sutter et al., 1994). CYP1B1 is expressed in a variety of humantumours, and can be inducible by TCDD in numerous cell types includingbreast, liver, lung, and kidney (see, e.g., Murray et al., 1997). CYP1B1is known to catalyse estradiol 4-hydroxylation metabolism. In untreatedculture, the constitutive rate of estradiol E₂ metabolism in MCF-7 cellsis minimal. However, treatment with TCDD causes a marked increase in therate of E₂ metabolism (see, e.g., Spink et al., 1994). Thus MCF-7 cellsin culture that are non-induced are metabolically analogous to normalcells that do not express CYP1B1, whilst TCDD-induced MCF-7 cellsexpress the CYP1B1 enzyme as is present in fresh human tumours.Therefore the cytotoxicity of compounds in non-induced MCF-7 cellscorrelates to the cytotoxicity of compounds against normal cells, whilstthe cytotoxicity of compounds against TCDD-induced MCF-7 cellscorrelates to the cytotoxicity of compounds against real tumours thatexpress CYP1B1. In this assay, a tumour selectivity factor greater than1 (and preferably greater than 1.5) is highly significant anddemonstrates that the compound has tumour selective cytotoxic activity.

The non-induced MCF-7 cell line is analogous enzymatically to normalcells that do not express catalytically active CYP1 family enzymes. Thecytotoxicity of compounds in non-induced MCF-7 cells correlates to thecytotoxicity of compounds against normal cells, whilst the cytotoxicityof compounds against TCDD-induced MCF-7 cells correlates to thecytotoxicity of compounds against real tumours that express CYP1B1.

Cells were maintained in RPMI 1640 with Glutamax 1 (Life Technologies)with 10% (v/v) heat inactivated foetal calf serum (Hybrimax. Sigma), at37° C., 5% CO₂/95% air with 100% humidity and passaged usingtrypsin/EDTA. 1×10³ cells were plated out in 100 μl medium per well of96-well flat-bottomed plates (Fisher). After 4 hours to allow adherance,100 μl of medium containing TCDD (British Greyhound Chromatography; 10μM stock in DMSO (dimethylsulfoxide)) or medium with 0.2% (v/v) DMSO ascontrol was added to each well to give a final concentration of 10 nMTCDD, 0.1% (v/v) DMSO, for 24 hours to allow maximal CYP expression. Themedium was then carefully aspirated and 100 μl fresh medium added.Within 30 minutes test compound was added in quadruplicate in 100 μlmedium (with or without inhibitors) at double the final concentrationfrom 100 mM stock in DMSO to give a final concentration of not more than0.1% (v/v) DMSO, or DMSO solvent alone at 0.1% (v/v) as control. Thecells were then allowed to grow on for 96 hours to give 80–90%confluence in the control wells. 50 μl MTT (Thiazol blue, Sigma) at 2mg/ml in Dulbecco's phosphate buffered saline-A, was then added to eachwell for 3 hours. All medium was aspirated, then the formazan productgenerated by viable cells was solubilized with 150 μl DMSO. Plates werevortexed and the absorbance at 540 nm determined using a plate reader.Results were expressed as the percentage of 100% (control) proliferationand the IC50 calculated using the line of best fit for a sigmoidal doseresponse curve with variable slope using Graphpad Prizm software. Alldeterminations were carried out in at least triplicate.

The selectivity differential factor (TSDF) is calculated by dividing theIC50 obtained from the MCF-10A data by the IC50 obtained from the MDA468data. A selectivity factor greater than 1 (and preferably greater than1.5) is highly significant and demonstrates that the compound has tumourselective cytotoxic activity.

The results of this assay are summarised in the table below. CompoundDMU-1.75 is 30-fold more toxic to “tumour” cells than to “normal” cells.Compound DMU-186 is 46-fold more toxic. Compound DMU-191 is 24-fold moretoxic. Compound DMU-103 (see compound 1 in Ikeda Shunichi et al., 1996)is 2-fold more toxic to normal cells than to cancer cells.

TABLE 1 Cytotoxicity TCDD-induced MCF-7 Cells MCF-7 Cells TumourSelectivity Compound IC50 (μM) IC50 (μM) Differential Factor DMU-174 2.00.6 3 DMU-175 15.0 0.5 30 DMU-176 20.0 1.1 18 DMU-185 8.0 0.66 12DMU-186 30.0 0.65 46 DMU-190 0.75 0.39 2 DMU-191 10.0 0.42 24 DMU-19220.0 1.6 12 DMU-188 4.5 0.6 8 DMU-189 8.3 2.5 3 DMU-401 6.8 0.8 9DMU-417 28 10 3 DMU-408 10 1.3 8 DMU-420 20 3.7 5 DMU-424 12 2 6 DMU-4320.7 0.08 9 DMU-103 0.04 0.08 0.5 DMU-402 6.2 6.2 1

FIG. 1 is a graph of cell survivial (%) versus concentration (μM) ofcompound DMU-175, for (A) the TCDD-induced MCF-7 cell line (▪) and (B)the MCF-7 cell line (▾).

This graph shows that compound DMU-175 has an IC50 of 15 μM inun-induced MCF-7 cells, but has an IC50 of 0.54 μM in TCDD-induced MCF-7cells. This illustrates a surprising and unexpected 28-fold increase inthe cytotoxic activity of DMU-175 by the induction of CYP1B1.Consequently, DMU-175 has a large therapeutic window; is active at muchlower doses; and will specifically target the tumour cells that expressCYP1B1, whilst normal cells will be preferentially spared.

MDA468 Tumour Cell Line Versus MCF-10A Normal Cell Line Assay

This cell culture based assay is performed using the two cell linesMDA-468 and MCF-10A. The MDA468 cell line is an advanced breast cancercell line, whilst the MCF-10A cell line is a normal breast cell line.

This assay was performed using the two cell lines MDA-468 and MCF-10Aaccording to the procedure described above for the MCF-7 assay, butwithout the addition of TCDD.

The tumour selectivity differential factor (TSDF) is calculated bydividing the IC50 obtained from the MCF-10A data by the IC50 obtainedfrom the MDA-468 data. In this assay, a tumour selectivity factorgreater than 1 (and preferably greater than 1.5) is highly significantand demonstrates that the compound has tumour selective cytotoxicactivity.

The results of this assay on Compound DMU-175, together with theclinically used anticancer agents tamoxifen, methotrexate, anddoxorubicin (adriamycin) for comparison, are summarised in the tablebelow. Surprisingly compound DMU-175 is 11-fold more toxic to cancercells than to normal cells. In contrast, the clinically used anticanceragent Doxorubicin is actually found to be 10-fold more toxic to normalcells than to cancer cells.

TABLE 2 Cytotoxicity MDA-468 MCF-10A (Breast Tumour) (Normal Breast)Tumour Selectivity Compound IC50 (uM) IC50 (uM) Differential FactorDMU-175 1.4 16 11 Tamoxifen 4.0 6.3 1.6 Methotrexate 0.04 0.06 1.5Doxorubicin 0.003 0.0003 0.1

FIG. 2 is a graph of cell survivial (%) versus concentration (μM) ofcompound DMU-175, for (A) the normal breast cell line MCF-10A (◯), and(B) the advanced breast cancer cell line MDA468 (●).

This graph shows that compound DMU-175 shows a low toxicity IC50 valueof 16 μM against the normal cell line, but a highly potent IC50 value of1.4 μM against the advanced tumour cell line. This illustrates asurprising and unexpected 11-fold tumour selectivity in the cytotoxicactivity of DMU-175.

Splenocyte Anti-Proliferation Assay

The splenocyte anti-proliferation assay has been developed to identifycompounds that have useful anti-inflammatory properties for thetreatment of auto-inflammatory diseases such as rheumatoid arthritis.See, for example, Yamashita et al., 1994. This well known assay isdescribed in detail in, for example, Mosmann, 1983. In this assay,splenocyte proliferation is stimulated by the inflammatory responseinducer conconavilin A (Con A). Cell proliferation is monitored bydetecting radiation (counts per minute, cpm) from a radio label(tritiated thymidine) which is incorporated only into proliferatingcells.

For example, compounds may be assayed as a solution in dimethylsulfoxide(DMSO) as solvent. A solvent control may also be tested for comparison.Other controls may be used. Compounds that exhibit anti-inflammatoryeffects at a concentration of less than 10 μM are considered to beuseful therapeutic agents.

The compounds of the present invention also show growth down-regulatoryeffects on splenocytes. Since splenocytes are involved in inflammation,these compounds are also useful as anti-inflammatory agents.

REFERENCES

A number of patents and publications are cited above in order to morefully describe and disclose the invention and the state of the art towhich the invention pertains. Full citations for these references areprovided below. Each of these references is incorporated herein byreference in its entirety into the present disclosure.

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1. A compound of the following formula:

wherein: R^(ALK) is primary or secondary aliphatic saturated C₂₋₆alkyl;two of R^(B2), R^(B3), R^(B4), and R^(B5) are —OMe; and the others areindependently —H or —OH; or: three of R^(B2), R^(B3), R^(B4), and R⁸⁵are —OMe: and the other is independently —H or —OH; each of R¹ and R² isindependently: —H, optionally substituted C₁₋₄alkyl, or optionallysubstituted C₅₋₂₀aryl; R^(A3) is —H, —OH, —OC(═O)R^(E), —OS(═O)₂OH, or—OP(═O)(OH)₂; R^(E) is: —H, optionally substituted C₁₋₆alkyl, optionallysubstituted C₃₋₂₀heterocyclyl, or optionally substituted C₅₋₂₀aryl; andpharmaceutically acceptable salt, solvates, amides, esters, and ethersthereof.
 2. A compound according to claim 1, wherein R^(ALK) is selectedfrom:


3. A compound according to claim 1, wherein R^(ALK) is selected from:-Et, -nPr, -iPr, -nBu, -iBu, -sBu, -nPe, and -nHex.
 4. A compoundaccording to claim 1, wherein R^(ALK) is selected from: -nPr, -nBu,-nPe, and -nHex.
 5. A compound according to claim 1, wherein: each of R¹and R² is independently —H or -Me.
 6. A compound according to claim 1,wherein: R¹ and R² are both —H.
 7. A compound according to claim 6,wherein: two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe; and theothers are independently —H or —OH.
 8. A compound according to claim 6,wherein: two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe; the othersare independently —H or —OH; and the two —OMe groups are not adjacent toeach other.
 9. A compound according to claim 6, wherein: two of R^(B2),R^(B3), R^(B4), and R^(B5) is —OMe; one of the others is —OH; and thelast is —H.
 10. A compound according to claim 6, wherein: two of R^(B2),R^(B3), R^(B4), and R^(B5) is —OMe; one of the others is —OH; the lastis —H; and the two —OMe groups are not adjacent to each other.
 11. Acompound according to claim 6, wherein: three of R^(B2) R^(B3) R^(B4)and R^(B5) is —OMe; and the others are independently —H or —OH.
 12. Acompound according to claim 6, wherein: two of R^(B2), R^(B3), R^(B4),and R^(B5) is —OMe; and the others are —H.
 13. A compound according toclaim 6, wherein: two of R^(B2), R^(B3), R^(B4), and R^(B5) is —OMe; theothers are —H; and the two —OMe groups are not adjacent to each other.14. A compound according to claim 6, wherein: three of R^(B2), R^(B3),R^(B4), and R^(B5) is —OMe; and the other is —H.
 15. A compoundaccording to claim 1, selected from compounds of the following formulae,and pharmaceutically acceptable salts, solvates, amides, esters, andethers thereof:


16. A compound according to claim 1, wherein: R^(E) is —CH₃, —CH₂CH₃,—C(CH₃)₃, or -Ph.
 17. A compound according to claim 1, wherein R^(A3) is—H.
 18. A compound according to claim 2, wherein R^(A3) is —H.
 19. Acompound according to claim 3, wherein R^(A3) is —H.
 20. A compoundaccording to claim 4, wherein R^(A3) is —H.
 21. A compound according toclaim 5, wherein R^(A3) is —H.
 22. A compound according to claim 6,wherein R^(A3) is —H.
 23. A compound according to claim 7, whereinR^(A3) is —H.
 24. A compound according to claim 8, wherein R^(A3) is —H.25. A compound according to claim 9, wherein R^(A3) is —H.
 26. Acompound according to claim 10, wherein R^(A3) is —H.
 27. A compoundaccording to claim 11, wherein R^(A3) is —H.
 28. A compound according toclaim 12, wherein R^(A3) is —H.
 29. A compound according to claim 13,wherein R^(A3) is —H.
 30. A compound according to claim 14, whereinR^(A3) is —H.
 31. A compound according to claim 15, wherein R^(A3) is—H.
 32. A compound according to claim 1, wherein R^(A3) is —OH.
 33. Acompound according to claim 2, wherein R^(A3) is —OH.
 34. A compoundaccording to claim 3, wherein R^(A3) is —OH.
 35. A compound according toclaim 4, wherein R^(A3) is —OH.
 36. A compound according to claim 5,wherein R^(A3) is —OH.
 37. A compound according to claim 6, whereinR^(A3) is —OH.
 38. A compound according to claim 7, wherein R^(A3) is—OH.
 39. A compound according to claim 8, wherein R^(A3) is —OH.
 40. Acompound according to claim 9, wherein R^(A3) is —OH.
 41. A compoundaccording to claim 10, wherein R^(A3) is —OH.
 42. A compound accordingto claim 11, wherein R^(A3) is —OH.
 43. A compound according to claim12, wherein R^(A3) is —OH.
 44. A compound according to claim 13, whereinR^(A3) is —OH.
 45. A compound according to claim 14, wherein R^(A3) is—OH.
 46. A compound according to claim 15, wherein R^(A3) is —OH.
 47. Acompound according to claim 1 selected from compounds of the followingformulae, and pharmaceutically acceptable salts, solvates, amides,esters, and ethers thereof:


48. A composition comprising a compound according to claim 1 and apharmaceutically acceptable carrier.
 49. A method of treating aproliferative condition in a patient comprising administering to saidpatient a therapeutically-effective amount of a compound according toclaim
 1. 50. A method according to claim 49, wherein the proliferativecondition is a solid tumour.
 51. A method according to claim 49, whereinthe proliferative condition is a solid tumour, and is a cancer of thelung, colon, breast, ovarian, prostate, liver, pancreas, brain, or skin.52. A method according to claim 49, wherein the proliferative conditionis a solid tumour, and is a cancer of the breast.
 53. A method accordingto claim 49, which is a method of prophylactically treating saidproliferative condition.
 54. A method of treating an inflammatorycondition in a patient comprising administering to said patient atherapeutically-effective amount of a compound according to claim
 1. 55.A method according to claim 54, wherein the inflammatory condition isrheumatoid arthritis, rheumatic fever, osteoarthritis, inflammatorybowel disease, psoriasis, or bronchial asthma.
 56. A method of diagnosisof a patient for the presence of tumour cells expressing the CYP1B1enzyme, comprising: (a) administering to the patient a compound asdefined below, wherein R^(A3) is —H; (b) determining the amount ofcorresponding hydroxylated metabolite, wherein R^(A3) is —OH, which issubsequently produced; and, (c) correlating the amount with the presenceor absence of the tumour cells in the patient; wherein the compound isselected from compounds of the following formula:

 wherein: R^(ALK) is primary or secondary aliphatic saturated C₂₋₆alkyl;two of R^(B2), R^(B3), R^(B4), and R^(B5) are —OMe; and the others areindependently —H or —OH; or: three of R^(B2), R^(B3), R^(B4), and R^(B5)are —OMe; and the other is independently —H or —OH; each of R¹ and R² isindependently: —H optionally substituted C₁₋₄alkyl, or optionallysubstituted C₅₋₂₀aryl; R^(A3) is —H; R^(E) is: —H optionally substitutedC₁₋₆alkyl, optionally substituted C₃₋₂₀heterocyclyl, or optionallysubstituted C₅₋₂₀aryl; and pharmaceutically acceptable salts, solvates,amides, esters, and ethers thereof.